Design of Novel Wound Dressing Composed of Collagen and Hyaluronic Acid Containing Epidermal Growth Factor

This research aims to develop a wound dressing composed of collagen (Col) and hyaluronic acid (HA) containing epidermal growth factor (EGF). First important issue is to contain EGF in the wound dressing in a stable state. The sheet-shaped sponge was manufactured by freeze-vacuum drying an aqueous solution of Col. Both sides of sponge were treated with ultraviolet (UV) irradiation to introduce intermolecular cross links between collagen molecules. This sponge was named Sponge-Col. Another sheet-shaped sponge was manufactured by freeze-vacuum drying an aqueous solution of HA containing EGF. This sponge was named Sponge-HA/EGF. The wound dressing was manufactured by laminating Sponge-Col on the top, Sponge-HA/EGF in the middle, and Sponge-Col on the bottom to create a sandwich structure. This method can prevent the reducing of EGF activity due to UV irradiation for intermolecular cross-linking. Second important issue is to enable gradual release of EGF from the wound dressing. The elution behavior of this wound dressing was investigated by measuring the weight change after immersion in water for a predetermined time. This wound dressing showed initially fast elution and subsequent very slow elution properties. The upper layer and lower layer Sponge-Col enabled gradual release of the middle layer Sponge-HA/EGF. This result suggests that EGF contained in the wound dressing is gradually released together with HA from the wound dressing. Third important issue is to provide moist wound-healing environment. The upper layer and lower layer Sponge-Col can provide the wound dressing with high water absorption and long-term water retention properties.

Process Control for Production of Human-like Collagen in Fed-batch Culture of Escherichia coli BL 21

Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5min and 4min intervals, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated. The studies showed that nitrogen source feeding in fast cycle could result in higher human-like collagen production than that in slow cycle; and the feedback regulation of glucose, increase of the pressure of fermentation bioreactor, and supply of oxygen-enriched air could all increase cell yield and human-like collagen production. The effects of inducement strength on protein expression were found important. When OD600 reached 90-100, the cultivation temperature was increased to 42℃ to begin induction for 2-3 h, and then shifted to 39℃ for 5-6h induction, the cell density and human-like collagen production could reach 96g·L-1 [DCW (dry cell mass)] and 19.8% (g·g-1 DCW) respectively.

Optimization of Fermentation Process for Human-like Collagen Production of Recombinant Escherichia coli Using Response Surface Methodology

In order to improve the production of human-like collagen III(HLC III)by fed-batch culture of recombinant Escherichia coli BL21,the Plackett-Burman and Box-Behnken design were applied to optimize the fermentation process parameters.Three variables(induction time,inoculum age and pH),which have significant effects on HLC III production,were selected from eight variables by Plackett-Burman design.With the regression coefficient analysis in the Box-Behnken design,a relationship between HLC III production and three significant factors was obtained,and the optimum levels of the three variables were as follows:induction time 3.2h,inoculum age 12.6 h and pH 6.7.The 3D response surface plots and 2D contour plots created by the Box-Behnken design showed that the interaction between induction time and pH and that between innoculum age and pH were significant.An average 9.68 g·L1HLC III production was attained in the validation experiment under optimized condition,which was 80%higher than the yield of 5.36 g·L1before optimization.

Analysis of Metabolic Products by Response Surface Methodology for Production of Human-like Collagen II

Recombinant Escherichia coli BL21 is used to produce human-like collagen. The key constituents of media are optimized using response surface methodology (RSM). Before thermal induction, the highest biomass production and the lowest production of some hazardous by-products, especially acetic acid, were obtained in the media containing 0.085 mol·L-1 glucose and 0.019 mol·L-1 nitrogen (carbon-nitrogen ratio, 4.47:1). After thermal induction, when the concentrations of glucose and nitrogen in the media were 0.065 mol·L-1 and 0.017 mol·L-1 , respectively (carbon-nitrogen ratio, 3.82:1), the productivity of human-like collagen per cell was the highest while that of acetic acid was the lowest. The extended analysis showed that the production of lactic acid and propionic acid increased while that of some intermediate acids of the tricarboxylic acid cycle decreased if the dose of glucose increased.

Beneficial effects of a novel shark-skin collagen dressing for the promotion of seawater immersion wound healing

Background: Wounded personnel who work at sea often encounter a plethora of difficulties. The most important of these difficulties is seawater immersion. Common medical dressings have little effect when the affected area is immersed in seawater, and only rarely dressings have been reported for the treatment of seawater-immersed wounds. The objective of this study is to develop a new dressing which should be suitable to prevent the wound from seawater immersion and to promote the wound healing.Methods: Shark skin collagen(SSC) was purified via ethanol de-sugaring and de-pigmentation and adjusted for p H. A shark skin collagen sponge(SSCS) was prepared by freeze-drying. SSCS was attached to an anti-seawater immersion polyurethane(PU) film(SSCS+PU) to compose a new dressing. The biochemical properties of SSC and physicochemical properties of SSCS were assessed by standard methods. The effects of SSCS and SSCS+PU on the healing of seawaterimmersed wounds were studied using a seawater immersion rat model. For the detection of SSCS effects on seawaterimmersed wounds, 12 SD rats, with four wounds created in each rat, were divided into four groups: the 3 rd day group, 5 th day group, 7 th day group and 12 th day group. In each group, six wounds were treated with SSCS, three wounds treated with chitosan served as the positive control, and three wounds treated with gauze served as the negative control. For the detection of the SSCS+PU effects on seawater-immersed wounds, 36 SD rats were divided into three groups: the gauze(GZ)+PU group, chitosan(CS)+PU group and SSCS+PU group, with 12 rats in each group, and two wounds in each rat. The wound sizes were measured to calculate the healing rate, and histomorphology and the immunohistochemistry of the CD31 and TGF-β expression levels in the wounded tissues were measured by standard methods.Results: The results of Ultraviolet-visible(UV-vis) spectrum, Fourier-transform infrared(FTIR) spectrum, circular dichroism(CD) spectra, sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE), and amino acid composition analyses of SSC demonstrated that SSC is type I collagen. SSCS had a homogeneous porous structure of approximately 200μm, porosity rate of 83.57%±2.64%, water vapor transmission ratio(WVTR) of 4500 g/m2, tensile strength of 1.79±0.41 N/mm, and elongation at break of 4.52%±0.01%. SSCS had significant beneficial effects on seawater-immersed wound healing. On the 3 rd day, the healing rates in the GZ negative control, CS positive control and SSCS rats were 13.94%±5.50%, 29.40%±1.10% and 47.24%±8.40%, respectively. SSCS also enhanced TGF-in the initial stage of the healing period. The SSCS+PU dressing effectively protected woundsβ and CD31 expression from seawater immersion for at least 4 h, and accelerated re-epithelialization, vascularization and granulation formation of seawater-immersed wounds in the earlier stages of wound healing, and as well as significantly promoted wound healing. The SSCS+PU dressing also enhanced expression of TGF-n and gauze dressings.β and CD31. The effects of SSCS and SSCS+PU were superior to those of both the chitosaConclusion: SSCS has significant positive effects on the promotion of seawater-immersed wound healing, and a SSCS+PU dressing effectively prevents seawater immersion, and significantly promotes seawater-immersed wound healing.

Kinetics of High Cell Density Fed-batch Culture of Recombinant Escherichia coli Producing Human-like Collagen

The kinetics of batch and fed-batch cultures of recombinant Escherichia coli producing human-like collagen was investigated. In the batch culture, a kinetic model of a simple growth-association system was concluded without consideration of cell endogeneous metabolism. The cell lag time, the maximum specific growth rate and Yx/s were determined as 1.75h, 0.65h^-1 and 0.51g·g^-1, respectively. In the fed-batch culture, different specific growth rates were set at （0.15, 0.2, 0.25h^-1） by the method of pseudo-exponential feeding, and the expressions for the specific rate of substrate consumption, the growth kinetics and the product formation kinetics of each phase were obtained. The result shows that the concentrations of cell and product can reach 77.5g·L^-1 and 10.2g·L^-1 respectively. The modal predictions are in good agreement with the experimental data.

Collagen/Chitosan Electrospun Nanofibrous Membrane for Wound Dressing

Collagen(Col)/chitosan(CS)nanofibrous membrane has great potential to be used as wound dressing.However,current Col/CS nanofibrous membrane produced from electrospinning can not offer sufficient mechanical strength for practical applications.Herein,a novel mixed solvent was used to prepare next-generation high-strength Col/CS nanofibrous membrane.Meanwhile,the optimal Col to CS weight ratio was investigated as well.The asproduced membrane was examined by scanning electron microscopy(SEM),attenuated total reflectance Fourier transform infrared spectroscopy(ATR-FTIR),differential scanning calorimetry(DSC),and XF-1A tester to study its morphological,chemical,thermal and mechanical properties.The preliminary results demonstrated that the mechanical properties of Col/CS nanofibrous membranes were enhanced substantially with the increase of CS weight ratios from 0 to 90%and the optimal Col to CS weight ratio was determined to be 1∶1.A promising way was presented to fabricate Col/CS electrospun nanofibrous membrane with sufficient mechanical strength for practical wound dressing applications.

Preliminary Clinical Study Using a Novel Wound Dressing Composed of Hyaluronic Acid and Collagen Containing EGF

This clinical trial aimed to evaluate the efficacy and safety of a novel wound dressing composed of hyaluronic acid (HA) and collagen (Col) containing epidermal growth factor (EGF), referred to as EGF-wound dressing. EGF-wound dressing was prepared by freeze-drying a mixed aqueous solution of high-molecular-weight HA, low-molecular-weight HA and heat-denatured Col containing EGF. EGF-wound dressing was applied to skin defects, such as intractable skin ulcers, burn ulcers, traumatic skin defects and skin donor-site wounds. The dressing was changed twice a week for a period of 6 weeks or longer, if necessary. The primary endpoints were size of wound area, formation of granulation tissue, extent of epithelialization, infection control and macroscopic appearance. Effectiveness, safety and overall clinical evaluation were scored by plastic surgeons, as authorized by the Japanese Society of Plastic and Reconstructive Surgery. This study was registered with the University Hospital Medical Information Network (UMIN0000005264). Healthy granulation tissue and rapid epithelialization were observed for a given period after application of EGF-wound dressing onto the wounds. Most cases were assessed as having achieved good or excellent results. This clinical study demonstrated that EGF-wound dressing was beneficial in the treatment of various skin defects.

Effect of calcium alginate dressing on the cytokine contents,collagen synthesis-degradation balance and apoptosis gene expression in the wound after perianal abscess surgery

Objective: To study the effect of calcium alginate dressing on the cytokine contents, collagen synthesis - degradation balance and apoptosis gene expression in the wound after perianal abscess surgery. Methods: Patients with perianal abscess who received surgical resection in the Eighth Hospital of Wuhan between May 2014 and February 2017 were selected and randomly divided into the group A who received calcium alginate dressing combined with kangfuxin solution and recombinant human epidermal growth factor for dressing change and the group B who received kangfuxin solution and recombinant human epidermal growth factor for dressing change. 3 d, 6 d and 9 d after dressing change, appropriate amount of wound tissue was collected to determine the expression of cytokines, collagen metabolites and apoptosis genes. Results: 3 d, 6 d and 9 d after dressing change, TGF-β1, Smad3, EGF and bFGF protein expression as well as Col-I, Col-II, Col-III, TIMP1 and TIMP2 protein expression in wounds of both groups of patients were increasing while Fas, FasL, Bax and Caspase-3 protein expression were decreasing, and TGF-β1, Smad3, EGF and bFGF protein expression as well as Col-I, Col-II, Col-III, TIMP1 and TIMP2 protein expression in wounds of group A were significantly higher than those of group B while Fas, FasL, Bax and Caspase-3 protein expression were significantly lower than those of group B. Conclusion: Calcium alginate dressing for wound dressing after perianal abscess surgery an increase the pro-proliferation cytokine expression, adjust the collagen synthesis - degradation balance and inhibit apoptosis, and it is conducive to wound healing.

Progress of Domestic Recombinant Human-Like Collagen Research and Application

By intercepting the most effective fragment of human collagen’s gene,recombinant human-like collagen is produced by genetic engineering technology and biotechnology.The collagen has good biocompatibility and can promote cell formation.Recombinant human-like collagen has great potential application value in medical biological materials,cosmetics and foods.In recent years,many specialized enterprises which are engaged in human-like collagen raw materials production have emerged.In this paper,a brief summary of the current recombinant human-like collagen raw material’s production and its latest research and development have been made.

强脉冲光联合类人胶原蛋白敷料治疗面部色素沉着的疗效及相关测评指标变化评析

目的:探究强脉冲光联合类人胶原蛋白敷料治疗面部色素沉着的疗效以及对VISIA系统色素沉着评分和表皮油脂量、TEWL的影响。方法:选取从2021年1月-2022年6月在笔者医院收治的100例女性面部色素沉着患者,将所有患者按随机数字表法分配为对照组(n=50)、观察组(n=50),对照组使用强脉冲光治疗,观察组在对照组的基础上联合类人胶原蛋白敷料治疗;比较两组患者的疗效、VISIA系统色素沉着评分、皮肤屏障功能:包括表皮含水量、表皮油脂含量和经皮水分丢失量(TEWL)、SAS、DLQI评分的变化情况和不良反应的发生情况。结果:观察组患者治疗后总有效率(88.00%,44/50)高于对照组(P<0.05);与治疗前相比,治疗后观察组和对照组的VISIA系统色素沉着评分、TEWL、SAS、DLQI评分显著降低,表皮含水量、表皮油脂含量显著升高(P<0.05),治疗后与对照组相比,观察组的VISIA系统色素沉着评分、TEWL、SAS、DLQI评分显著降低,表皮含水量、表皮油脂含量显著升高(P<0.05);不良反应发生率,观察组和对照组比较,差异无统计学意义(P>0.05)。结论:强脉冲光联合类人胶原蛋白敷料治疗面部色素沉着的效果显著,可以促进色素沉着消退,改善皮肤屏障功能,改善患者生活质量和心理状态,且安全性较高。

凯氏定氮法测定医用重组胶原蛋白修复敷料中蛋白质

建立了测定医用重组胶原蛋白修复敷料中胶原蛋白含量的凯氏定氮法。参照《中华人民共和国药典》2020年版通则0731蛋白质含量测定方法第一法,将钨酸沉淀法进行了改良,对样品进行处理得到非蛋白氮样品溶液,用全自动凯氏定氮仪测定样品与非蛋白氮样品中的氮含量,并计算样品中的胶原蛋白含量。样品中非蛋白氮的平均质量分数为0.0100%,相对标准偏差为1.60%(n=6);胶原蛋白的平均质量分数为0.0999%,相对标准偏差为2.13%(n=6),样品加标平均回收率为98.82%。该方法可以有效消除样品中非蛋白氮的干扰,准确测定医用重组胶原蛋白修复敷料的胶原蛋白含量。

昆海牡甲汤辅助微创旋切术对乳腺良性肿块切除术的疗效

目的:探究昆海牡甲汤辅助微创旋切术联合重组人源胶原蛋白功能敷料对乳腺良性肿块切除术患者创面止血及凝血功能的影响。方法:选取2019年9月-2021年9月我院收治的乳腺良性肿块患者90例,所有患者均接受乳腺良性肿块麦默通微创旋切术治疗,按照患者的治疗意愿分为观察组和对照组各45例,其中对照组患者术后给予重组人源胶原蛋白功能敷料,观察组患者在对照组基础上给予昆海牡甲汤口服,其他止血及治疗措施与观察组一致,观察记录两组患者的术中出血情况、住院成本及效果比、治疗前后血常规及凝血功能情况。结果:观察组患者的术中出血量、术后出血量、血肿发生率均显著低于对照组,差异有统计学意义(P <0.05);两组患者的住院时间对比未见显著差异(P>0.05),观察组患者的住院成本高于对照组患者(P <0.05),但观察组的有效率及成本效果比显著优于对照组(P <0.05);术前及术后,观察组患者的白细胞计数(WBC)、红细胞计数(RBC)、血红蛋白(HGB)水平均显著低于对照组,血小板(PLT)水平显著高于对照组(P <0.05);两组患者术前及术后凝血功能指标比较,差异无统计学意义(P>0.05)。结论:重组人源胶原蛋白功能敷料联合昆海牡甲汤对乳腺良性肿块切除术患者创面止血具有良好的效果,不会影响患者的凝血功能,具有较好的安全性和治疗成本效果比。

浅谈Ⅱ类重组胶原蛋白创面敷料产品注册监管的思考

重组胶原蛋白为原料的Ⅱ类重组胶原蛋白创面敷料被广泛用于医疗领域。目前国家药监局及国家医疗器械技术审评中心颁布的关于重组胶原蛋白原料及Ⅱ类重组胶原蛋白创面敷料的相关标准及指导原则远不能满足Ⅱ类重组胶原蛋白敷料注册审评、生产质量控制及监管的需求。文章针对Ⅱ类重组胶原蛋白创面敷料注册审评、质量控制及监管等一些问题进行了思考及分析,并提出了建议,以期为医疗器械监管部门的决策提供一定参考。

重组胶原蛋白的性能及在创面敷料中的应用

文章首先综述了重组胶原蛋白在创面敷料中的应用,重点关注其独特的性能特点及其在不同类型创面治疗中的作用。重组胶原蛋白因其优异的生物相容性、低免疫原性和可控降解性,成为创面敷料领域的理想生物材料。文章详细讨论了重组胶原蛋白的定义、命名规则、性能特点,以及其在液体、凝胶和冻干粉等不同形式敷料中的应用,其次,探讨了重组胶原蛋白在烧伤治疗、创面愈合和皮肤修复中的创新应用,以及相关的质量控制措施;最后,总结了重组胶原蛋白敷料的优势,并展望了未来的发展方向。

胶原贴敷料对面部寻常痤疮患者皮肤屏障功能及局部免疫的影响

目的探究胶原贴敷料对面部寻常痤疮患者皮肤屏障功能及局部免疫的影响。方法将2020年6月至2022年11月江南大学附属医院的148例面部寻常痤疮患者根据随机数表法分为对照组(74例)和观察组(74例)。对照组进行红蓝光照射治疗,观察组则在对照组的基础上加用胶原贴敷料。比较两组面部寻常痤疮治疗总有效率、治疗前后的痤疮相关指标(皮损数量、皮损E值及M值)、皮肤屏障功能指标[角质层含水量(SCH)及经皮水分丢失(TEWL)]及局部免疫指标(CD3^(+)、CD4^(+)及CD8^(+))。结果观察组的面部寻常痤疮治疗总有效率为94.59%,高于对照组的82.43%,差异有统计学意义(P<0.05);治疗前两组的痤疮相关指标、皮肤屏障功能指标及局部免疫指标比较,差异无统计学意义(P>0.05),治疗2、4周后观察组的上述方面均改善,观察组的痤疮相关指标、局部免疫指标及TEWL低于对照组,而SCH则高于对照组,差异有统计学意义(P<0.05)。结论胶原贴敷料在面部寻常痤疮患者中的应用效果较好,有助于患者皮肤屏障功能及局部免疫状态的改善。

重组胶原蛋白液体敷料鉴别检验方法适用性研究

重组胶原蛋白敷料是目前市场较火的一种皮肤护理类产品,可通过在创面形成保护层,起物理屏障作用。用于非慢性创面(如浅表性创面、激光/光子/果酸换肤/微整形术后创面)的护理,为创面愈合提供微环境。对敷贴类产品中所含的重组胶原蛋白鉴别的检测,是确保产品质量可靠的指标之一。本文研究了一款重组胶原蛋白液体敷料的鉴别方法的适用性。分别使用肽图法、N末端氨基酸序列法和等电聚焦电泳法对该试验样品中的重组胶原蛋白进行鉴别,检测结果发现肽图法、N末端氨基酸序列法对该样品具有适用性。而用现有的等电聚焦电泳法未发现有条带检出,该方法不适用。企业在选择重组胶原蛋白鉴别方法时应结合产品实际情况,对鉴别方法进行研究,采用适宜的方法。

Recent progresses of collagen dressings for chronic skin wound healing

The skin plays a fundamental role in regulating the body's internal balance and protecting against external traumas.A broad variety of environmental risk factors frequently result in acute skin wounds,whose inappropriate treatments would lead to chronic skin wounds that are difficult to heal.Traditional dressings have been widely used to repair chronic skin wounds,however their drawbacks such as insufficient hemostatic efficacy and non-moist environment have severely limited their clinical applications.As the principal component of skin,collagen has always been a research hotspot in the field of chronic skin wounds due to its advantages of low antigenicity,high biocompatibility and superior bioactivity.Collagen-based dressings have been increasingly developed to heal the chronic wounds during the past decades,arising from their capability in decreasing protein and electrolyte losses in wound exudate,preventing bacterial contamination,permitting less painful dressing changes,and improving the healing quality.This review overviews recent progress of collagen dressings for chronic skin wound healing.Various commonly used wound dressings for wound management have been first introduced.Collagen wound dressings have been categorized as films,sponges,hydrogels,nanofibers,and powders,and their efficacy has been compared.The critical functions of collagen dressings in wound healing,such as stopping bleeding,shortening inflammation,promoting angiogenesis,and stimulating tissue regeneration have been elaborated.The clinical applications of collagen dressings to repair different types of chronic wounds have been thoroughly summarized.A comprehensive list of commercialized collagen dressings has been updated,and an outlook of collagen dressings have been finally speculated.

Design of Medical Devices Using Spongy Sheet Composed of Hyaluronic Acid and Collagen

This research aims to obtain useful information for development of medical devices such as wound dressing and tissue anti-adhesive product, using a spongy sheet composed of hyaluronic acid (HA) and collagen (Col). The spongy sheets were manufactured by freeze vacuum drying of HA and Col aqueous solution, followed by UV irradiation to introduce intermolecular crosslinks between Col molecules. These spongy sheets are referred to as Sponge-A (ratio of HA/Col = 5/1) and Sponge-B (ratio of HA/Col = 5/5). Both surfaces of Sponge-A and Sponge-B treated with UV irradiation for 15 minutes are referred to as Sponge-A-15 and Sponge-B-15, respectively. The weight change of spongy sheet was determined by immersing a peace of spongy sheet in water at 37°C. The weight of sponge-A-15 collected 1/2, 1, 3, 7 days after immersion in water was 63.5%, 62.1%, 56.6%, 54.4% of the original weight, respectively. The weight of Sponge-B-15 was 78.3%, 76.7%, 79.1%, 71.9% of the original weight, respectively. The weight change of spongy sheet was determined by immersing a peace of spongy sheet in water containing collagenase at 37°C. The weight of Sponge-A-15 collected 6, 8, 10, 12 hours after immersion in water containing collagenase (0.0005 w/v%) was 65.7%, 59.8%, 57.9%, 55.2% of the original weight, respectively. The weight of Sponge-B-15 was 63.5%, 52.1%, 42.0%, 43.2% of the original weight, respectively. This spongy sheet is considered to have the unique structure, where HA molecules are entrapped in an intermolecular cross-linked network structure of Col molecules. When immersed in water containing collagenase, the weight loss of spongy sheet is accelerated by easy extraction of HA molecules from the enzymatic degraded Col network structure. The performance of wound dressing and tissue anti-adhesive product is considered to depend on appropriate ratio of HA and Col, and also on appropriate rate of intermolecular crosslinks between Col molecules. These findings obtained in this study provide useful information for product development such as wound dressing and tissue anti-adhesive product.

Design of Collagen-Based Sponge Device for Use in Oral Surgery

This study aims to develop various types of collagen devices for use in oral surgery. The targets are the sheet-shaped sponges to cover mucosal defect wounds (product-1) and gingival defect wounds (product-2) and the cylindrical sponge to fill tooth extraction sockets (product-3). The sheet-shaped sponges were manufactured by freeze-vacuum drying the aqueous solution of collagen (Col) and heat-denatured collagen (Col’) at a composition ratio of 2/1, 1/1 or 1/2. Both surfaces of the sheet-shaped sponge were treated by ultraviolet (UV) irradiation for 5, 10 or 15 minutes to introduce intermolecular crosslinks between collagen molecules. The elution behavior of each collagen sponge was investigated by immersing the sponge in water for a predetermined time and then by measuring the weight change. The collagen sponge composed of Col and Col’ with a composition ratio of 2/1 that was treated by UV irradiation for 15 minutes showed very slow elution properties. This sheet-shaped sponge is the top candidate for product-1. The collagen sponge composed of Col and Col’ with a composition ratio of 2/1 that was treated by UV irradiation for 5 minutes showed slightly fast elution properties. This sheet-shaped sponge is the top candidate for product-2. Next, the cylindrical sponge was manufactured by freeze-vacuum drying the aqueous solution of Col and Col’ at a composition ratio of 2/1. Both sides of the cylindrical sponge were treated by UV irradiation for 15 minutes. This sponge showed initially fast elution properties and subsequent very slow elution properties. This cylindrical sponge is a good candidate for product-3. As a basic design, the sheet-shaped sponge and the cylindrical sponge should be biodegraded and absorbed by the time new tissue formation is completed.