[화학공학] 히알루론산의 구조, 성능, 개질 및 응용에 대한 자료입니다

루 론 산 (hyaluronan, hyaluronic acid, HA) is a glycosaminoglycan that occurs naturally in living organisms. It was first isolated from the vitreous humor of cattle in 1934 by Karl Meyer and John Palmer of Columbia University in the United States. They named it “hyaluronic acid”, which comes from the words “hyalo-oid” and “uronic acid” [1]. Later, Endre Balazs coined the term “hy aluronan” in 1986 to name hyaluronic acid in line with the international naming convention for polysaccharides, to cover various molecular forms (including acid and salt forms) [2]. Hyaluronic acid is an important component of the cell matrix and various tissues, and has a variety of important physiological functions, such as regulating cell proliferation, migration and differentiation; natural moisturizing; lubricating joints to protect cartilage; regulating protein synthesis; regulating inflammatory responses; regulating immune function; promoting wound healing, etc.

루 론 acid's의 독특한 점탄성, 생체 적합성 및 분해성으로 인해 안과 수술 보조제, 수술 후 유착 방지제, 상처 치유 및 재생 보조제, 약물 운반체, 조직 공학 스캐폴드 등으로 포함하여 생의학 분야에 광범위하게 응용되고 있습니다.본 글에서는 히알루론산의 구조, 성질 및 화학적 개질 방법에 대해 설명하고, 현재 생의학 분야에서의 응용 현황에 대해 설명한다.루 론 acid'의 독특한 구조적 특성 및 우수한 특성은 생물 의학 분야에서 매우 유망한 응용 분야를 가지고 있음을 의미합니다.이 리뷰의 목적은 researchers&를 높이는 것입니다#39;히알루론산에 대한 종합적인 설명을 제공함으로써 히알루론산에 대한 관심을 갖게하고, 새로운 히알루론산 생의학 소재 설계를 위한 몇 가지 지침을 제공하고자 한다.

히알루론산의 구조, 성질 및 생리적 기능 1

1. 히알루론산의 화학구조 1

Hyaluronic acid is a member of the glycosaminoglycan (also known as mucopolysaccharide) family. Like other glycosaminoglycans, hyaluronic acid is a high-molecular-weight linear polysaccharide composed of repeating disaccharide units of aminohexose and hexuronic acid. However, it is the only non-sulfated glycosaminoglycan and the only glycosaminoglycan that is not covalently linked to nuclear proteins to form proteoglycans. Unlike most glycosaminoglycans, hyaluronic acid is synthesized on the cell membrane via membrane proteins, rather than via the cell's 골기구 [3].자연 루 론의 disaccharide 단위 산은 D-glucuronic 산의 그리고 N-acetyl-D-glucosamine로 구성 된, β-1으로 연결 되어 있는, 3 glycosidic 본드, 그리고 disaccharide 장치는 β-1으로 연결 되어 있고, 4 glycosidic 본드, i. e. [(1 → 3)-β-D-GlcNAc-(1 → 4)-β-D-GlcUA] (그림 1 참조), 분자의 무게를 싣고 가고 10월 7일까지 Da [4].모두 설탕을 채택 β-configuration, 수산기들과, 카르, acetamido고 e-bonding 위치에서 hydroxymethyl 그룹, 루 론 산은 매우 안정적인 열정적으로 만들고 있다.

1. 히알루론산의 2가지 성질

Hyaluronic acid is a white, amorphous solid with no smell. It is highly hygroscopic, soluble in water but insoluble in organic solvents. 이hydrophilic groups in the molecular structure of hyaluronic acid are all in the parallel positions of the sugar rings, while the hydrophobic hydrogen atoms form a hydrophobic region in the axial direction. Due to the hydrogen bonding between the monosaccharide molecules in the molecular chain, the hyaluronic acid molecular chain forms a rigid columnar helical structure in space. In an aqueous solution, the hyaluronic acid molecules form an expanded, random coil structure. At lower concentrations, these hyaluronic acid chains also entangle with each other to form a continuous three-dimensional network structure with unique rheological properties. Water molecules are fixed in the network formed by hyaluronic acid molecules through hydrogen bonds and are not easily lost. Studies have shown that hyaluronic acid can adsorb about 1000 times its own weight in water, making it the best natural water-retaining substance found in nature. A 1% solution can form a gel, but it is easily fluid under pressure and can pass through the narrow passage of an injection needle. It is a pseudoplastic material. The extraordinary rheological properties of hyaluronic acid solutions make them ideal lubricants, capable of separating the surfaces of most tissues and allowing them to slide along each other.

1. 히알루론산의 분해 3

The degradation of hyaluronic acid in the body can be seen as a depolymerization process in which glycosidic bonds break, mainly through enzymatic hydrolysis and free radical degradation. The enzymatic degradation of hyaluronic acid in the body is mainly carried out by the hyaluronidase family, which has six members: HYAL-1, HYAL-2, HYAL-3, HYAL-4, HYAL-P1 and PH-20 [5]. Among them, the two most active enzymes are HYAL-1 and HYAL-2. HYAL-2 (located on the cell membrane) cleaves high molecular weight HA (>1MDa) into fragments of 20kDa. HYAL-1 (located in lysosomes) then cleaves these fragments into tetroses, which are further converted into monosaccharides by the action of other enzymes (e.g. β-glucuronidase, β-N-acetylglucosaminidase). Since these degradation products are natural substances that are present in the human body, they can participate in the body's 자신의 제거 과정.반면 조직염증 등으로 생성된 활성산소는 글리코시드 결합을 쪼개어 히알루론산의 산화분해를 일으키기도 한다.히알루론산의 이화작용은 현장 (예를 들어, 세포 밖 기질에서), 세포 내, 림프절에서 일어난다.사슬이 긴 히알루론산은 효소와 활성산소에 의해 현장에서 분해되어 더 작은 히알루론산 올리고당을 생성합니다.이후 이러한 올리고당류는 세포와 림프절 내부에서 추가적인 대사과정을 거쳐 최종적으로 순환계로 들어가 간과 신장에 의해 제거된다 [6].

1. 히알루론산의 생리기능 4

Hyaluronic acid is an important component of the extracellular matrix. In the past, hyaluronic acid was considered to be a simple space-filling substance, and it was only gradually that its importance was recognized. Due to its high water absorption, the primary role of hyaluronic acid in the human body is structural support and moisture retention. It provides lubrication and shock absorption for cells and other extracellular matrix components (including collagen and elastin), while regulating the water balance of tissues and providing a favorable environment for cell migration and proliferation. Hyaluronic acid also has a large number of negatively charged carboxyl groups on its backbone, which act as ion exchangers and can regulate the concentration of cations around cells. In addition, hyaluronic acid also acts as a signaling molecule, participating in cell signaling and regulating various cell activities, including cell proliferation, migration, differentiation, and adhesion, by binding to various protein receptors on the extracellular matrix and cell membrane. Thus, it plays a role in regulating physiological functions of the body, For example, hyaluronic acid can promote the aggregation of white blood cells at the site of inflammation through binding to the CD44 receptor, thereby promoting the body's 면역 항염증 효과 [7].

This signal-regulating effect of hyaluronic acid is related to its molecular weight, with hyaluronic acids of different molecular weights triggering different signal pathways. High molecular weight hyaluronic acid exhibits anti-angiogenic, scar-inhibiting and anti-inflammatory effects, while low molecular weight hyaluronic acid (<100kDa) exhibits the opposite effects, promoting inflammation, immune stimulation, scar formation and angiogenesis [8]. The cause of this difference is still uncertain. One hypothesis is that 고 분자량 히알루론산 has the effect of aggregating receptor proteins on cell membranes, while low molecular weight hyaluronic acid does not have this effect [9], thus causing differences in receptor activity and resulting in different physiological functions.

Hyaluronic acid is an intelligent moisturizing factor that can adjust its water absorption according to the relative humidity of the surrounding environment, regulating the water balance of cells and tissues. In the skin, these highly moisturizing hyaluronic acids form an extracellular colloidal matrix with a high water content together with collagen and elastin, giving the skin resilience and elasticity. At the same time, hyaluronic acid also has the effect of scavenging free radicals. As mentioned above, free radicals can oxidize and degrade hyaluronic acid, and hyaluronic acid uses this degradation reaction to remove free radicals in the body through its own rapid metabolism.

히알루론산은 활액의 주성분이기도 하며, 높은 점탄성은 관절을 보호하는 데 중요한 역할을 합니다.그것은 조직 사이의 마찰을 감소시키는 보행과 같은 낮은 충격 주파수에서 점성 액체입니다;스트레스의 충격을 완충하는 가동과 같은 높은 충격 주파수의 탄성 액체;그리고 하중을 받으면 젤 같은 탄성체가, 관절에 가해지는 압력을 줄여주는 쿠션 역할을 한다 [10].

Hyaluronic acid also plays a role in promoting tissue wound healing and is a recognized major compound in this process. It plays an important role in the activation and regulation of immune responses, the promotion of angiogenesis, and cell proliferation and migration. During the inflammatory phase, high molecular weight hyaluronic acid increases, absorbing water to expand and produce a porous scaffold suitable for cell migration, inhibiting the migration of neutrophils, and reducing the inflammatory response. During the proliferation phase, hyaluronan oligosaccharides promote angiogenesis and the migration of fibroblasts to the wound tissue, where they construct a new extracellular matrix. During the reconstruction phase, hyaluronic acid regulates scar formation [11].

히알루론산 2 공업 생산

Hyaluronic acid is widely found in the cell matrix and lubricating fluid of various tissues in animals, including human umbilical cords, joint synovial fluid, skin, thoracic lymphatic fluid, vitreous humor, and rooster combs. The rooster comb is currently the animal tissue found to have the highest hyaluronic acid content (see Table 1) [4]. The extraction process of hyaluronic acid generally involves a complete set of processes such as homogenization, extraction, precipitation, and impurity removal of these freshly collected hyaluronic acid-rich tissues to finally obtain hyaluronic acid with high purity. Although the extraction method has a simple process flow, it is restricted by the limited source of raw materials, low efficiency, and high cost, and has gradually been replaced by the fermentation method.

The use of microbial fermentation to prepare hyaluronic acid first appeared in the 1970s, but it was not until 1985 that Shiseido in Japan first reported the use of Streptococcus fermentation to produce hyaluronic acid. This led to the development of the biological fermentation method, which gradually replaced the traditional animal tissue extraction method and has become the mainstream international method of hyaluronic acid production today [12]. Currently, the commercially produced hyaluronic acid strains include Streptococcus and Bacillus subtilis.

히알루론산의 화학적 변형 3

…의 거주 시간순수한 히알루론산인체에서는 피부나 관절에 주사 후 반감기가 24시간 이내로 비교적 짧다 [13].이 때문에 생의학 분야에서 응용이 크게 제한된다.그러나 히알루론산은 탈아세틸화에 의해 노출된 카르복실기, 히드록실기, 아미노기를 포함한 여러 활성 그룹으로 인해 더욱 화학적으로 변형될 수 있으며, 이를 통해 더 나은 기계적 강도, 레올로지 특성 및 효소 가수분해에 대한 저항성 등을 가질 수 있어 생의학적 응용의 범위가 확대될 수 있다.

3.1 카복실기 개질

3.1.1 아미 화 반응

The carboxyl group of hyaluronic acid can be activated by carbodiimides, 2-chloro-4,6-dimethyl-1,3,5-triazine (CDMT), 2-chloro-1-methyliodopyridine (CMPI), 1, 1,1'-카보닐디이미다졸 (CDI) 등이 활성화 [14~17] 된 후 효율적으로 아미노화합물과 반응하여 아미드 결합을 형성한다 (그림 2 참조). 그 중에서 가장 널리 사용되는 활성제는 1-에틸-3-(3-dimethylaminopropyl) 카보디이미드 (EDC)이다.반응 메커니즘은 다음과 같습니다:먼저 EDC의 활성화 된 카르복실기는 o-아세틸 isourea 중간체를 형성 한 다음 아미노기는 친핵성 공격을 수행하여 아미드 결합을 형성합니다.또한 o-아세틸 이소레아 중간체는 물과 반응하여 안정적인 부산물인 n-아세틸루레아를 형성하기 위해 빠르게 재배열되기 쉬우므로, n-아세틸루레아의 형성을 막기 위해 활성화 중에 N-succinimide (NHS)나 하이드록시벤조트리아졸 (HOBT)을 첨가하여 안정적이고 가수분해물에 강한 중간체를 형성한다 (그림 3 참조) [18].

On the other hand, the optimum pH for the EDC activation reaction is 3.5~4.5, and amino groups have a high pK a value. Under these pH conditions, the nucleophilicity of protonated amino groups is reduced, and their reactivity with activated carboxyl groups is also reduced. Replacing amino groups with hydrazides with a low pK a (pK a ≈2~3) can increase the reactivity [19]. Hyaluronic acid gels prepared with dihydrazides as cross-linking agents have stronger mechanical properties. When an excess of adipoyl dihydrazide (ADH) is used to react with hyaluronic acid, only a monofunctionalization reaction occurs, forming a stable hyaluronic acid-ADH derivative that retains the other hydrazide as a reaction site for further functionalization (see Figure 2). In practice, many hyaluronic acid-drug precursors are formed by the reaction of the hyaluronic acid-hydrazide intermediate with imine-activated drugs rather than by the reaction of hyaluronic acid itself, because the main by-product when hyaluronic acid is used directly is N-acetylurea.

3. 1. 2 Esterification

In addition to reacting with amino compounds, the carboxyl group of hyaluronic acid can also undergo an esterification reaction with fatty or aromatic alcohols. The above-mentioned activating reagents can also be used to catalyze the esterification of the hyaluronic acid carboxyl group. The activated hyaluronic acid can also undergo a crosslinking reaction with its own hydroxyl groups to form a self-crosslinking gel (the crosslinking structure does not contain a crosslinking agent). In addition to esterification with alcohols, hyaluronic acid can also react with haloalkanes and epoxides to form ester bonds (see Figure 4) [20 , 21] .

3. 2 히드록시기 개질

3. 2. 1 Etherification

Due to the presence of hyaluronidase, the half-life of natural hyaluronic acid in the human body is relatively short. Therefore, various hyaluronic acid fillers on the market generally use chemical cross-linking to improve their resistance to enzymatic hydrolysis and prolong their retention time in the body. In 1964, Laurent et al. [22] first reported the cross-linking reaction of hyaluronic acid. They used 1,2,3,4-diepoxbutane as the cross-linking agent, and the reaction occurred under strong alkaline conditions with a pH of 13-14. Currently, the cross-linking agents used by major manufacturers around the world include 1,4-butanediol diglycidyl ether (BDDE), 1,2,7,8-diepoxyoctane (DEO), divinyl sulfone (DVS), etc. (see Figure 5) [23] , are mainly used to cross-link hyaluronic acid through etherification. Etherification generally takes place under strongly alkaline conditions. Here, the hydroxyl groups undergo deprotonation (pK a ≈ 10) to form strongly nucleophilic oxygen anions, which preferentially add nucleophilically to deprotonated carboxyl groups to form ether bonds. Under acidic conditions (pH 2~4. 5), the deprotonation of the hydroxyl group is reduced, and the ester bond is mainly formed by the attack of the negatively charged carboxyl group on the epoxy group (see Figure 5) [24]. However, Tomihata and Ikada [25] found that under weak acidic and neutral conditions (pH = 4.7, 6.1, 8.0), the product is still dominated by ethers.

3. 2. 2 Esterification

The 히알루론산의 히드록시기 can also undergo esterification reactions with activated carboxylic acids, anhydrides, and active groups such as acid chlorides. For example, Coradini et al. [26] reported the use of butyric anhydride to react with the hydroxyl group on the trimethylpyridine salt of hyaluronic acid in the presence of pyridine or dimethylaminopyridine to form a hyaluronic acid-butyric acid precursor. This hyaluronic acid-butyric acid precursor drug not only retains the original pharmacological effects of butyric acid, but also promotes the uptake of butyric acid by cells and improves the effect of butyric acid in inhibiting the growth of tumor cells. In fact, hyaluronic acid-butyric acid is completely endocytosed into MCF-7 human breast cancer cells under the mediation of the CD44 receptor, showing relatively obvious tumor targeting.

3. 2. 3기타 반응

The hydroxyl groups of hyaluronic acid글루타알데히드와 교차 연결 반응을 일으켜 헤미칼릭스 [26]를 형성하는 등 다른 반응을 겪을 수도 있다.이 반응은 알데히드 그룹을 활성화하고 반응을 촉매하기 위해 산성 조건을 필요로 한다.그러나 결과물인 hemiketal은 산성 조건에서 가수분해되기 쉬우므로 [27] 교반된 생성물을 안정화시키기 위해 반응 말기에 중화가 필요하다.또한, 히알루론산의 히드록시기 역시 시아노겐 브로마이드에 의해 활성화되어 수용액 단계에서 아민 화합물과 반응하여 카바메이트를 형성할 수 있다 [28].

3. 3 탈 아세틸화 및 아미 화

The free amino group formed by deacetylation of the acetyl group on hyaluronic acid can also be used as an active site for modification reactions. It can react with activated carboxylic acids to form amide compounds, or even undergo self-crosslinking with its own carboxyl group to form a gel. However, deacetylation, even under mild conditions, can cause degradation of hyaluronic acid [29], so this method is generally not used for hyaluronic acid modification.

3. 4   복잡 한 수정

Hyaluronic acid can also be used in combination with other materials to take advantage of their respective advantages and compensate for their deficiencies. For example, hyaluronic acid and chitosan can be combined to form nanoparticles through electrostatic interaction, which can be used to load papain and form new surfactants[30]; hyaluronic acid and gelatin can be combined through emulsification-coagulation, and smooth, wrinkled and porous microspheres can be obtained by different post-treatment methods [31]; the combination of hyaluronic acid and hydroxypropyl methyl cellulose can improve the resistance of the gel to enzymatic hydrolysis [32, 33]; the combination of hyaluronic acid and collagen will give it better mechanical properties.

3. 5 금속 복합체

Hyaluronic acid is rich in O and N atoms, and can form coordination bonds with a variety of metal ions, such as Fe3+, Zn2+, Cu2+, Ni2+, etc. Coordination changes the structure of hyaluronic acid in solution and gives it more biological functions[34]. For example, Curiosin gel from Gedeon Richter is a complex of hyaluronic acid and Zn2+, which changes the structure of hyaluronic acid from a random coil to a spherical structure through coordination, reducing the thickness of the bound water molecule layer and making the bond more stable. Clinical trials have shown that this gel can effectively promote wound healing and prevent wound infection.

히알루론산 및 유도체의 생의학적 응용 4

루 론 acid's unique properties make it suitable for a wide range of biomedical applications. Balazs [35] divides the clinical applications of hyaluronic acid and its derivatives into five categories.

(1) 점안술:연약한 조직을 보호하고 안과 수술 등 외과 수술에 공간을 제공;

(2) 점증술:피부, 괄약근, 성대, 인두조직 등 조직 공간을 채우고 확장하는 것;

(3) 점분리:유착 및 과도한 흉터를 방지하기 위해 수술이나 외상으로 인한 손상된 결합 조직 표면을 분리;

(4) 점성보충 (viscosupplimentation):관절염에서 윤활액을 교체하는 등 조직액을 대체하거나 보충하여 통증을 완화시키는 것;

(5) 점성 보호 (viscoprotection):건강하거나 손상된 조직 표면을 건조하거나 유해한 환경의 영향으로부터 보호하고, 조직 표면 치유를 촉진한다.

4. 1 안과

Hyaluronic acid is a major component of the eye's 유리체 유머이며 주로 안과 수술에서 백내장 수술이나 안구 내 수정체 삽입과 같은 수술 중 손실된 유리체 유머를 대체하기 위해 사용됩니다.한편, 히알루론산은 안과 수술에서 점탄성 보호제로도 사용되며 각막 상피를 보호하고 기계적 충격을 완충하며 안구 전방의 적절한 깊이와 모양을 유지하고 안구 내 조직을 보호하며 유리체 탈출증을 막고 외과적 수술을 용이하게 한다 [36].히알루론산은 안구건조증 치료를 위한 안약의 주성분이기도하다.눈물막 파열 시간을 효과적으로 연장하고 안구건조증 환자의 눈깜빡임 횟수를 줄이며 건조증, 자극, 가려움, 통증 등의 증상을 완화시킬 수 있다.

4. 피부 필러 2가지

Hyaluronic acid is a natural moisturizer that is widely found in skin tissue, and its concentration can reach 2. 5g/L. As we age, the amount of hyaluronic acid in the skin gradually decreases, leading to dehydration of the dermis, deepening of wrinkles, and loss of elasticity. Hyaluronic acid is widely used as a skin filler to treat facial aging due to its high viscoelasticity, plasticity, biodegradability, good biocompatibility, and lack of species specificity. According to statistics from the International Society of Aesthetic Plastic Surgery (ISAPS), the number of cases in which hyaluronic acid fillers are used ranks second among minimally invasive cosmetic treatments, after botulinum toxin. However, the natural hyaluronic acid in the human body has a very short maintenance cycle and cannot guarantee the long-term effect of filling and modification. Therefore, physical or chemical cross-linking protection methods are generally used to increase the resistance of hyaluronic acid to enzymatic hydrolysis and prolong its retention time in the body.

4. 유착 방지 및 상처 치유 3

Postoperative tissue adhesion is a major problem in surgical procedures, which can lead to serious long-term clinical complications, affecting the results of surgical procedures and causing pain and inconvenience to patients. A large number of studies have shown that hyaluronic acid plays an important role in preventing adhesion and promoting wound healing. The mechanism of hyaluronic acid in preventing tissue adhesion mainly includes: (1) separating tissues through physical shielding, which can also shield inflammatory mediators and bacteria, thus playing a protective role; (2) promoting the dissolution of blood fibrin, while stimulating the expression of CD44 receptors to promote the proliferation of mesenchymal cells; (3) enhancing the function and activity of macrophages, regulating collagen synthesis, reducing the deposition of blood fibrin, promoting wound healing and reducing scar formation; (4) forming a protective film on the tissue surface to reduce mechanical damage and provide lubrication and moisture; (5) absorbing and expanding to compress bleeding points and suppress bleeding[38] .

Epidermal growth factor (EGF), fibroblast growth factor (bFGF), etc. are now widely used in skin wound repair, but these products are generally in the form of freeze-dried powder, which needs to be stored in the refrigerator before use, and has a very short half-life, so it needs to be applied repeatedly every day. Yamamoto et al. [39] reported a dual-layer wound dressing formed by high-molecular-weight and low-molecular-weight hyaluronic acid, where the high-molecular-weight cross-linked hyaluronic acid forms the upper layer of the supplement and the low-molecular-weight hyaluronic acid, arginine, vitamin C derivatives and EGF form the lower layer. Experimental results show that this wound supplement can maintain the activity of EGF and promote the release of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF).

4. 4 관절염

Hyaluronic acid is the main component of articular cartilage and synovial fluid. In normal, healthy joints, movement can be carried out almost friction-free and pain-free. However, when joint diseases such as osteoarthritis or rheumatoid arthritis occur, the concentration of hyaluronic acid in the synovial fluid decreases significantly, the molecular weight decreases significantly, and the cartilage is also degraded and destroyed, causing the joint movement to become stiff and pain due to bone-on-bone friction. Injecting exogenous high molecular weight hyaluronic acid into the joint restores the synovial fluid to a normal state and promotes the gradual natural repair of cartilage. At the same time, the injected hyaluronic acid also improves the biological environment of the joint cavity, promotes the synthesis of endogenous hyaluronic acid, and improves joint function. However, because the half-life of hyaluronic acid in the body is short, repeated and frequent injections are required for the treatment of joint lesions, which increases the patient' s 고통을 받고 있다.최근 조던 등 [40]은 히알루론산과 키토산을 혼합해 만든 새로운 형태의 겔을 발표했다.키토산을 첨가하면 히알루론산의 분해 방지 능력이 향상될 뿐만 아니라 치료 효과도 향상된다.이 연구는 관절 질환 치료를 위한 점성 히알루론산 보충제를 개선할 수 있는 새로운 방향을 제시한다.

In recent years, there has been some debate about whether this viscoelastic supplement therapy is effective in treating arthritis. The second edition of the “Evidence-Based Guidelines for the Treatment of Knee Osteoarthritis” issued by the American Academy of Orthopaedic Surgeons in 2013 clearly states that hyaluronic acid is not recommended for the treatment of symptomatic knee osteoarthritis. They believe that although many studies have shown that the effect of high molecular weight hyaluronic acid on the treatment of osteoarthritis is statistically different compared to the control, this difference does not meet the minimum clinically important difference (MCII) standard and therefore does not have a clinically significant difference.

4. 5 마약 운반체 (Drug carrier)

Hyaluronic acid has the potential to be used as a drug carrier due to its good biocompatibility, high hydrophilicity, high viscoelasticity, degradability and specific binding to cell surface receptors (such as CD44 and RHAMM). On the other hand, from the chemical structure of hyaluronic acid, it has multiple reaction sites, including carboxyl groups, hydroxyl groups and acetylated amino groups, which can be used to construct drug precursors and carriers using a variety of chemical modification methods. At present, hyaluronic acid and its derivatives have been used to construct drug delivery systems for a variety of drugs, including anti-inflammatory drugs, anti-tumor drugs, protein peptide drugs and gene drugs, which can significantly prolong the blood circulation residence time of drugs, increase cellular uptake, improve bioavailability, reduce the amount of drug administered, and reduce adverse reactions [8, 41]. Zhong et al. [42] reported a reduction-sensitive, reversibly cross-linked hyaluronic acid nanoparticle composed of a hyaluronic acid-lysine-lipoic acid (HA-Lys-LA) covalent bond, which is cross-linked by a disulfide bond under the catalysis of 1, 4-dithio-D,L-threitol (DTT) catalysis, the drug doxorubicin (DOX) is cross-linked by disulfide bonds to improve the residence time of the drug under physiological conditions.

This nanocarrier specifically binds to the CD44 receptor overexpressed on the surface of MCF-7 human breast cancer cells resistant to DOX through hyaluronic acid located on the surface, thereby increasing the cellular uptake of the drug. The nanocarrier then swells and releases the drug by catalysing the breakage of the disulfide bond by glutathione, which is overexpressed in tumor cells, effectively inhibiting tumor growth (as shown in Figure 6). Park et al. [43] also reported the use of a similar drug carrier for siRNA transfection. They designed and synthesized a hyaluronic acid-poly(dimethylaminoethyl methacrylate) (HPD) grafted polymer as a siRNA delivery vehicle, and cross-linked it via a disulfide bond. In vitro experiments have shown that the cross-linked siRNA complex (C-siRNA-HPD) is more stable and can be more effectively taken up by melanoma cells overexpressing CD44, thereby improving siRNA transfection efficiency. In vivo experiments have shown that after systemic administration in mice, C-siRNA-HPD selectively accumulates in tumors, demonstrating its tumor targeting properties.

4. 조직공학 6

Tissue engineering is a new interdisciplinary subject that emerged in the 1980s and has become a research hotspot in tissue and organ regeneration medicine in recent years. Hyaluronic acid is an important component of many tissues in the human body and is a major component of the extracellular matrix. It affects cell proliferation, migration and differentiation, and promotes wound healing, making it an ideal raw material for tissue engineering. However, the weak mechanical properties, high swelling properties, smooth surface structure and lack of resistance to enzymatic hydrolysis of hyaluronic acid gels also limit their application in tissue engineering. Therefore, in order to improve the possibility of using them as scaffolds for tissue engineering, necessary chemical modifications are required to compensate for their deficiencies. One good method is to select other biomaterials for compounding, which can combine the advantages of multiple materials to complement each other' s 단점.예를 들어, 알긴산나트륨과 히알루론산이 교반되어 다공성 합성 겔을 형성할 수 있다.고분자의 농도와 두 다당류의 비율을 조절함으로써 합성 겔의 팽창 속도, 공극률 및 효소 가수분해에 대한 저항성을 조절할 수 있으며, 세포 부착 및 증식에 좋은 생물학적 환경을 제공할 수 있다 [44].

4. 7 Biomimetics

High-throughput screening of drugs is generally done through 2D in vitro cytological evaluation, but this method differs greatly from the actual results in vivo. Using 3D scaffolds to simulate the cell microenvironment is more in line with actual conditions. Hyaluronic acid is an important component of the extracellular matrix, and using it to construct a 3D culture medium will be more suitable for mimicking the in vivo growth environment of cells. Hyaluronic acid itself is negatively charged, which hinders cell adhesion, so it needs to be combined with other biomaterials to promote cell adhesion. Zhang et al. [45] used hyaluronic acid and chitosan to construct a 3D porous scaffold material for mimicking the extracellular matrix microenvironment of U-118MG human malignant glioma cells as a 3D culture medium for high-throughput screening of anti-tumor drugs. Compared with 2D culture medium, hyaluronic acid-chitosan scaffold culture medium can promote the formation of tumor spheroids and upregulate the expression of CD44, nestin, Musashi-1, GFAP and HIF-1α proteins.

5 결론

Hyaluronic acid has a history of more than 60 years since it was first used in human medicine in the late 1950s. Due to its special rheological properties and physiological functions, hyaluronic acid is widely used in the biomedical field. So far, the research on the design of new hyaluronic acid derivatives has made great progress, and more and more hyaluronic acid products have been developed to fill the gaps in biomedical applications. This paper reviews the structural properties, synthetic modification and biomedical applications of hyaluronic acid. However, there are still many unanswered questions about the physiological functions of hyaluronic acid. At present, commercially available hyaluronic acid biomaterials still have certain defects that require further improvement to promote the wider application of hyaluronic acid in the biomedical field.

참조

[1] 마이어 K, 팔머 JW다.  J Biol Chem, 1934년, 107: 629~634.

[2] 발라즈에아, 로랑 TC, Jeanloz RW,다.  바이오켐제이 (Biochem J, 1986년, 235: 903~903.

[3]Weigel PH, 하스콜 VC, 지난 밤에 M.  J Biol Chem, 1997년, 272: 13997~14000.

[4] 코간 G, 용매 L, 스턴 R, Gemeiner P.  바이오 테크놀 렛 (Biotechnol Lett), 2007년, 29일: 17~25.

[5]Csoka AB, Frost기, 스턴 R다.  매트릭스 바이올 (Matrix Biol), 2001년, 20: 499~508.

[6] 데 볼레 K, 글로가우 R, 코노 T, 네이선 M, 테젤 A, 로카-마르티네즈 J-X, 팔리왈 S, Stroumpoulis D에 있다.  피부과 Surg, 2013년, 39대:1758~1766.

[7] 테르메르 C, 썰맨 JP, 사이 먼 JC다.  Trends Immunol, 2003년, 24: 112~114.

[8]Schante CE, 주버 G, Herlin C, Vandamme TF다.  Carbohydr Polym, 2011년, 85: 469~489.

[9] 지앙 D, 리앙 J, 팬 J, 유 S (Yu S), 첸 S, Luo Y, 프레스트위치 GD, Mascarenhas 음, Garg HG, 퀸다, 호머 RJ, 골드스타인 박사, 부칼라 R, 이 PJ, 메드지토프 R, PW 숭고하다.  Nat Med, 2005, 11: 1173년~1179.

[10] 링 페이슈, 허연리, 장청.식품의약품안전처, 2005, 7:1~3.

[11] 진옌, 리다웨이, 주메이화, 천장잉.식품과 마약, 2014, 16:373~376.

[12] Cui Yuan, Duan Qian, Li Yanhui.장춘이공대학 (자연과학편), 2011, 34:101~106.

[13] 갈색 TJ, 로랑 UBG, 프레이저 JRE다.  Exp 생리술, 1991년, 76: 125~134.

[14] 다니셰프스키 1세, Siskovic E.  카보히닥터레스 (Carbohydr Res), 1971년, 16: 199~205.

[15] 마그나니 A, 라푸올리 R, 람포니 S, Barbucci R.  Polym Adv Technol, 2000년, 11: 488~495.

[16] 버그만 K, 엘빙슨 C, 힐본 J, 스벤스크 G, 보우 덴은 T.  Biomacromolecules, 2007년, 8: 2190~2195.

[17] 벨리니 D, Topai A.  WO2000001733A1다. 2000.

[18] 불피트 P, Aeschlimann D에 있다.  JBiomed Mater Res, (영어) 1999년, 47: 152~169.

[19]Pouyani T, Prestwich GD다.  생물공액화학 (Bioconjugate Chem), 1994년, 5: 339~347.

[20] 펠레 티어 S, 휴버트 P, 라피크 F, 파얀 E, Dellacherie E.  Carbohydr Polym, 2000년, 43: 343~349.

[21] 벤처리프사, 스리니바산 A, 호케이 F, 홀링거 조, Matyjaszewski K, Washburn NR다.  생체, 2008년, 29일: 1739~1749.

[22] 로랑 TC, 헬싱 K, Gelotte B다.  Acta Chemica Scandinavica 스칸디나비카, 1964년, 18: 274~275.

[23] 전지안, 루이지리.cn 102321258b.2012.

[24] 드벨더안, Malson T.  US4886787A다. 1985.

[25] 토미하타 K, Ikada Y.  생체, 1997년, 18: 189~195.

[26] 코라디니 D, 펠리자로 C, Miglierini G, 다이돈 MG, Perbellini A.  정수 J 암, 1999년, 81: 411~416.

[27] 콜린스 MN, Birkinshaw C.  J Appl Polym Sci, 2007년, 104: 3183~3191.

[28]Mlcochova P, 바이스트리키 S, 슈타이너 B, 마초바 E, Koos M, 벨브니 V, Krcmar M.  Biopolymers, 2006년, 82: 74~79

[29] 크레센지 V, 프란체스카 안젤리 A, 씨그레 알 (Segre AL), 카피타니 D, 만니나 L, 레니에 D, 벨리니 D에 있다.  마크로몰 바이오 sci, 2002년, 2: 272~279.

[30]Zhao D, Wei W, Zhu Y, Sun J, Hu Q, Liu X.  Macromol Biosci, 2015, 15:558~567.

[31]Zhou Z, He S, Huang T, Peng C, Zhou H, Liu Q, Zeng W, Liu L, Huang H, Xiang L, Yan H.  Polym Bull, 2015, 72:713~723.

[32] 지안준, 리루이지.CN 102492180B.2014.

[33] 지안준, 리루이지.CN 102911380A.2013.

[34] 진옌, 링 페이슈, 장천민.중국 생화학 약물 저널, 2008, 29427-429.

[35] Garg HG, Hales CA. Hyaluronan의 Chemistry and Biology, UK:Elsevier, 2004, 415-455.

[36] 장레이, 우디, 선웨이, 선중데.한국미생물학회지, 2006, 26:100-103.

[37] 범서치, 조우진성, 천규하.해협 약국, 2003, 15:252.

[38] 링 페이슈, 관후아시.중국약학저널, 2005, 40:1527-1530.

[39] 야마모토 A, 시미즈 N, Kuroyanagi Y.  J  Artif 장기, 2013년, 16: 489~494.

[40] 카데를리 S, 불로처 C, 필렛 E, Watrelot-Virieux D, 루게몽 알 (Rougemont AL), 로저 T, Viguier E, Gurny R, 스카포자 L, 조던 O다.  인트 제이 팔 (Int J Pharm), 2015년, 483: 158~168.

[41] 장위, 연퀴 ' e.화학 진보, 2006, 18:1684~1690.

[42] Zhong Y, Zhang J, Cheng R, Deng C, Meng F, Xie F, Zhong z.j 제어 해제, 2015, 205:144-154.

[43] Yoon HY, Kim HR, Saravanakumar G, Heo R, Chae SY, Um W, Kim K, Kwon I C, Lee JY, Lee DS, Park JC, Park JH.  J 제어 해제, 2013, 172:653~661.

[44]Chen Y H, Li J, Hao Y B, Qi J X, Dong N G, Wu C L, Wang Q. J Appl Polym Sci, 2015, 132:41898.

[45] Florczyk SJ, Wang K, Jana S, Wood DL, Sytsma SK, Sham JG, Klevit FM, Zhang M. Biomaterials, 2013, 34:10143-10150.