HAN Hongshuai, SONG Mizhao, LI Jiaxin, et al. Extraction of Collagen and Preparation and Characterization of Nanofibers[J]. Science and Technology of Food Industry, 2023, 44(19): 182−190. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022100267.
Citation: HAN Hongshuai, SONG Mizhao, LI Jiaxin, et al. Extraction of Collagen and Preparation and Characterization of Nanofibers[J]. Science and Technology of Food Industry, 2023, 44(19): 182−190. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022100267.

Extraction of Collagen and Preparation and Characterization of Nanofibers

More Information
  • Received Date: October 27, 2022
  • Available Online: August 04, 2023
  • In order to improve the extraction rate and utilization rate of collagen in sheepskin, sheepskin collagen was extracted by acid enzyme compounding method, and collagen-based nanofibers were prepared by electrospinning technology. Taking the extraction rate of sheepskin collagen as the evaluation index, the effects of four factors, namely material-liquid ratio, acetic acid concentration, pepsin concentration and enzymatic hydrolysis time, on the extraction effect of sheepskin collagen were investigated, and the optimal level of single factors were determined. On this basis, the process conditions of sheepskin collagen extraction were optimized by orthogonal experimental design. The effects of enzymatic hydrolysis on the structural properties of collagen were explored by biochemical techniques such as ultraviolet spectrum scanning, infrared spectrum scanning, SDS-PAGE mapping and scanning electron microscopy. Collagen and polylactic acid were then compounded and electrospun to obtain collagen-based nanofibers. The results showed that the best process for extracting sheepskin collagen by acid enzyme compounding method was as follows: Material-liquid ratio of 1:25 g/mL, acetic acid concentration of 1.2 mol/L, pepsin dosage of 1.0%, enzymatic hydrolysis time of 72 h. Under these conditions, the extraction rate of sheepskin collagen was 38.42%±0.49%. Ultraviolet spectroscopy showed that sheepskin collagen had a maximum UV absorption peak around 230 nm. Infrared spectrum scanning and SDS-PAGE map analysis showed that sheepskin collagen was mainly composed of α1, α2 and β three subunit components, which belonged to type I collagen, and the spatial structure of collagen was intact. Scanning electron microscopy showed that the fiber network structure of sheepskin collagen was relatively intact. The collagen-based nanofibers obtained by electrospinning had a diameter of 418.02±183.77 nm, a tensile strength of 3.616±0.386 MPa, and an elongation at break of 8.69%±1.95%, which had excellent physical properties and the potential to be used as a cell scaffold. This study would provide a theoretical basis for improving the high-value utilization of sheepskin and sheepskin collagen, and also provide theoretical support for the development of collagen-based nanomedical fiber products.
  • [1]
    周丹, 沈艳琴, 武海良, 等. 胶原蛋白的应用研究现状[J]. 合成纤维,2022,51(8):5−8. [ZHOU D, SHEN Y Q, WU H L, et al. Application and research status of collagen[J]. Synthetic Fibers,2022,51(8):5−8.

    ZHOU D, SHEN Y Q, WU H L, et al. Application and research status of collagen[J]. Synthetic Fibers, 2022, 51(8): 5-8.
    [2]
    ZHANG M, ZHAO D, ZHU S R, et al. Overheating induced structural changes of type I collagen and impaired the protein digestibility[J]. Food Research International,2020,134(6):1−9.
    [3]
    郑少东, 崔国廉, 胡杨, 等. 动物皮在创伤修复及止血材料中的应用[J]. 中国皮革,2014,43(1):42−45. [ZHENG S D, CUI G L, HU Y, et al. Application of animal skin in wound repair and hemostatic materials[J]. China Leather,2014,43(1):42−45.

    ZHENG S D, CUI G L, HU Y, et al. Application of animal skin in wound repair and hemostatic materials[J]. China Leather, 2014, 43(1): 42-45.
    [4]
    金月华, 曹佳雷, 孙霞, 等. 我国皮革服装行业的质量状况及改进策略[J]. 北京皮革,2020(6):16−20. [JIN Y H, CAO J L, SUN X, et al. Quality status and improvement strategy of leather garment industry in China[J]. Beijing Leather,2020(6):16−20.

    JIN Y H, CAO J L, SUN X, et al. Quality status and improvement strategy of leather garment industry in China[J]. Beijing Leather, 2020(6): 16-20.
    [5]
    任蕙铭, 李明娟. 黑龙江省畜牧产品营销现状及创新方式的思考[J]. 黑龙江畜牧兽医,2020,602(14):29−31. [REN H M, LI M J. Thinking on the marketing status and innovation methods of animal husbandry products in Heilongjiang Province[J]. Heilongjiang Animal Husbandry and Veterinary Medicine,2020,602(14):29−31.

    REN H M, LI M J. Thinking on the marketing status and innovation methods of animal husbandry products in Heilongjiang Province[J]. Heilongjiang Animal Husbandry and Veterinary Medicine, 2020, 602(14): 29-31.
    [6]
    高金龙. 山羊皮中胶原蛋白的提取及理化特性研究[D]. 呼和浩特: 内蒙古农业大学, 2010.

    GAO J L. Extraction and physicochemical properties of collagen in goat skin[D]. Hohhot: Inner Mongolia Agricultural University, 2010.
    [7]
    SAFIYA N, CASPARUS J R, MARK C L, et al. Collagen extraction from various waste bovine hide sources[J]. Waste and Biomass Valorization,2020,11(11):5687−5698. doi: 10.1007/s12649-019-00843-2
    [8]
    JAFARI H, LISTA A, SIEKAPEN M M, et al. Fish collagen: Extraction, characterization, and applications for biomaterials engineering[J]. Polymers,2020,12(10):2230−3340. doi: 10.3390/polym12102230
    [9]
    JIA A B, LI M, KANG L Z, et al. Fabrication and comprehensive characterization of biomimetic extracellular matrix electrospun scaffold for vascular tissue engineering applications[J]. Journal of Materials Science,2019,54(15):10871−10883. doi: 10.1007/s10853-019-03667-6
    [10]
    BEHESHTKHOO N, KOUHBANANI M, DEHGHANI F, et al. Fabrication and properties of collagen and polyurethane polymeric nanofibers using electrospinning technique for tissue engineering applications[J]. Journal of Environmental Treatment Techniques,2019,7(4):802−807.
    [11]
    KIM D, LEE M, YANG J, et al. Dual skin-whitening and anti-wrinkle function of low-molecular-weight fish collagen[J]. Journal of Medicinal Food,2022,25(2):192−204. doi: 10.1089/jmf.2021.K.0124
    [12]
    CHATTERJEE N, SUKUMARAN H, DARA P, et al. Nano-encapsulation of curcumin in fish collagen grafted succinyl chitosan hydrogel accelerates wound healing process in experimental rats[J]. Food Hydrocolloids for Health,2022,52(2):1−8.
    [13]
    MALLICK M, ARE R, BABU A. An overview of collagen/bioceramic and synthetic collagen for bone tissue engineering[J]. Materialia,2022,131(1):391−492.
    [14]
    黄艳萍, 但年华, 但卫华. 静电纺丝制备胶原基复合纳米医用纤维的研究进展[J]. 材料导报,2019,33(19):3322−3327. [HUANG Y P, DAN N H, DAN W H. Research progress on preparation of collagen-based composite nanomedical fibers by electrospinning[J]. Materials Reports,2019,33(19):3322−3327.

    HUANG Y P, DAN N H, DAN W H. Research progress on preparation of collagen-based composite nanomedical fibers by electrospinning [J]. Materials Reports, 2019, 33(19): 3322-3327.
    [15]
    SHEIK H, FARNA Z, HORRAM K, et al. Preparation, characterization, and surface modification of polycaprolactone-based nanofibrous scaffold by grafting with collagen for skin tissue engineering[J]. Regenerative Engineering and Translational Medicine,2022,8(4):545−562. doi: 10.1007/s40883-022-00254-1
    [16]
    WANG F, ZHOU M, JIA Q. Evaluation of dynamic mechanical and cytotoxic properties of electrospun poly (lactic acid)/cellulose nanocrystalline composite membranes[J]. Journal of Physics: Conference Series,2021,1759(1):31−41.
    [17]
    张鹏. 聚乳酸电纺纳米纤维结构性能研究与应用拓展[D]. 南昌: 东华理工大学, 2015.

    ZHANG P. Research and application development of structural properties of polylactic acid electrospun nanofiber[D]. Nanchang: East China University of Technology, 2015.
    [18]
    林琳, 牛一帆, 史振霞, 等. 猪皮中胶原蛋白的提取[J]. 廊坊师范学院学报(自然科学版),2020,20(1):58−61. [LIN L, NIU Y F, SHI Z X, et al. Extraction of collagen from pig skin[J]. Journal of Langfang Normal University (Natural Science Edition),2020,20(1):58−61.

    LIN L, NIU Y F, SHI Z X, et al. Extraction of collagen from pig skin[J]. Journal of Langfang Normal University (Natural Science Edition), 2020, 20(1): 58-61.
    [19]
    周瑞, 陈舜胜. 鱼皮胶原蛋白提取方法研究及其在食品中的应用[J]. 上海农业学报,2021,37(1):129−135. [ZHOU R, CHEN S S. Research on extraction method of fish skin collagen and its application in food[J]. Shanghai Journal of Agricultural Sciences,2021,37(1):129−135.

    ZHOU R, CHEN S S. Research on extraction method of fish skin collagen and its application in food[J]. Shanghai Journal of Agricultural Sciences, 2021, 37(1): 129-135.
    [20]
    王杉杉. 牦牛皮胶原蛋白的提取及其活性肽的制备[D]. 绵阳: 西南科技大学, 2018.

    WANG S S. Extraction of yak hide collagen and preparation of its active peptides[D]. Mianyang: Southwest University of Science and Technology, 2018.
    [21]
    王彦珍, 宋秘钊, 王娟, 等. 聚乙烯醇羊皮水解物复合纳米纤维制备与表征[J]. 棉纺织技术,2021,49(9):11−14. [WANG Y Z, SONG M Z, WANG J, et al. Preparation and characterization of polyvinyl alcohol sheepskin hydrolysate composite nanofibers[J]. Cotton Textile Technology,2021,49(9):11−14.

    WANG Y Z, SONG M Z, WANG J, et al. Preparation and characterization of polyvinyl alcohol sheepskin hydrolysate composite nanofibers[J]. Cotton Textile Technology, 2021, 49(9): 11-14.
    [22]
    黄可承, 宫萱, 唐嘉诚, 等. 水产品副产物胶原蛋白制备方法及应用[J]. 精细化工,2022,39(9):1757−1766. [HUANG K C, GONG X, TANG J C, et al. Preparation method and application of aquatic product by-product collagen[J]. Fine Chemicals,2022,39(9):1757−1766.

    HUANG K C, GONG X, TANG J C, et al. Preparation method and application of aquatic product by-product collagen[J]. Fine Chemicals, 2022, 39(9): 1757-1766.
    [23]
    张晓亮. 兔皮胶原蛋白的提取及抗菌敷料的制备[D]. 天津: 天津工业大学, 2020.

    ZHANG X L. Extraction of rabbit skin collagen and preparation of antibacterial dressing[D]. Tianjin: Tianjin Polytechnic University, 2020.
    [24]
    任戈一, 马雪莲, 潘丽, 等. 不同提取工艺下牛骨胶原蛋白的结构特性[J]. 食品工业科技,2022,43(14):27−33. [REN G Y, MA X L, PAN L, et al. Structural characteristics of bovine bone collagen under different extraction processes[J]. Science and Technology of Food Industry,2022,43(14):27−33.

    REN G Y, MA X L, PAN L, et al. Structural characteristics of bovine bone collagen under different extraction processes[J]. Science and Technology of Food Industry, 2022, 43(14): 27-33.
    [25]
    MONTALBANO G, TOMASINA C, FIORILLI S, et al. Biomimetic scaffolds obtained by electrospinning of collagen-based materials: Strategies to hinder the protein denaturation[J]. Materials,2021,14(16):4360−4370. doi: 10.3390/ma14164360
    [26]
    IFFA A F, PRIHARTINI W, Muhammad A, et al. Poly-L-lactic acid (PLLA)-chitosan-collagen electrospun tube for vascular graft application[J]. Journal of Functional Biomaterials,2018,9(2):32−34. doi: 10.3390/jfb9020032
    [27]
    王彦珍, 宋秘钊, 王娟, 等. 羊皮水解蛋白/聚乙烯醇纳米纤维静电纺丝影响因素[J]. 毛纺科技,2021,49(10):6−10. [WANG Y Z, SONG M Z, WANG J, et al. Influencing factors of electrospinning of parchment hydrolyzed protein/polyvinyl alcohol nanofibers[J]. Wool Spinning Science and Technology,2021,49(10):6−10.

    WANG Y Z, SONG M Z, WANG J, et al. Influencing factors of electrospinning of parchment hydrolyzed protein/polyvinyl alcohol nanofibers[J]. Wool Spinning Science and Technology, 2021, 49(10): 6-10.
    [28]
    李萍, 张慧君, 郭浩, 等. 糖基化改性玉米醇溶蛋白膜的性能及硬胶囊体外释放分析[J]. 农业工程学报,2021,37(14):302−309. [LI P, ZHANG H J, GUO H, et al. Performance of glycosylated modified zein membrane and in vitro release analysis of hard capsules[J]. Transactions of the Chinese Society of Agricultural Engineering,2021,37(14):302−309.

    LI P, ZHANG H J, GUO H, et al. Performance of glycosylated modified zein membrane and in vitro release analysis of hard capsules[J]. Transactions of the Chinese Society of Agricultural Engineering, 2021, 37(14): 302-309.
    [29]
    陈曼, 何明, 郭妍婷, 等. 静电纺丝蛋白纳米纤维膜的改性研究进展[J]. 化工新型材料,2017,45(7):21−23. [CHEN M, HE M, GUO Y T, et al. Research progress on modification of electrospinning protein nanofiber membranes[J]. New Chemical Materials,2017,45(7):21−23.

    CHEN M, HE M, GUO Y T, et al. Research progress on modification of electrospinning protein nanofiber membranes[J]. New Chemical Materials, 2017, 45(7): 21-23.
    [30]
    单钱艺, 沈岳明, 张明兴, 等. 正交试验优化超声波-复合酶水解法提取中华鳖油的工艺[J]. 食品工业科技,2022,43(22):256−264. [SHAN Q Y, SHEN Y M, ZHANG M X, et al. Orthogonal test to optimize the process of extracting Chinese turtle oil by ultrasonic-complex enzymatic hydrolysis[J]. Science and Technology of Food Industry,2022,43(22):256−264.

    SHAN Q Y, SHEN Y M, ZHANG M X, et al. Orthogonal test to optimize the process of extracting Chinese turtle oil by ultrasonic-complex enzymatic hydrolysis[J]. Science and Technology of Food Industry, 2022, 43(22): 256-264.
    [31]
    杨璇. 基于超声酸酶法和模拟肠消化制备鱼皮胶原蛋白肽及肽钙螯合物研究[D]. 镇江: 江苏大学, 2020.

    YANG X. Preparation of fish skin collagen peptide and peptide calcium chelate based on ultrasonic acid enzyme method and simulated intestinal digestion[D]. Zhenjiang: Jiangsu University, 2020.
    [32]
    甘文梅, 张炜, 田格, 等. 响应面法优化牦牛骨胶原蛋白的提取工艺研究[J]. 化学研究与应用,2020,32(9):1535−1540. [GAN W M, ZHANG W, TIAN G, et al. Study on optimization of yak bone collagen extraction process by response surface method[J]. Chemical Research and Application,2020,32(9):1535−1540.

    GAN W M, ZHANG W, TIAN G, et al. Study on optimization of yak bone collagen extraction process by response surface method[J]. Chemical Research and Application, 2020, 32(9): 1535-1540.
    [33]
    JOSUE J V, EDUARDO S M. Collagen and elastin scaffold by electrospinning for skin tissue engineering applications[J]. Journal of Materials Research,2019,34(16):2819−2827. doi: 10.1557/jmr.2019.233
    [34]
    程国清. 静电纺载药胶原蛋白纳米纤维膜的制备及性能研究[D]. 天津: 天津工业大学, 2017.

    CHENG G Q. Study on the preparation and properties of electrospun-loaded collagen nanofiber membranes[D]. Tianjin: Tianjin Polytechnic University, 2017.
    [35]
    BAKIR G, GIROUARD B E, WIENS R, et al. Orientation matters: polarization dependent IR spectroscopy of collagen from intact tendon down to the single fibril level[J]. Molecules,2020,25(18):4285−4295. doi: 10.3390/molecules25184285
    [36]
    LUO X S, GUO Z Z, HE P, et al. Study on structure, mechanical property and cell cytocompatibility of electrospun collagen nanofibers crosslinked by common agents[J]. International Journal of Biological Macromolecules,2018,113:476−486. doi: 10.1016/j.ijbiomac.2018.01.179
    [37]
    RANI C, SAFIRA A, SURYADININGRAT M, et al. Characterization of tilapia collagen-loaded chitosan nanofibers synthesized by electrospinning method for wound dressing[J]. IOP Conference Series:Earth and Environmental Science,2022,1036(1):1−12.
    [38]
    吴国良, 任天斌, 曹春红, 等. 静电纺丝法制备PLCL/明胶与PLCL/胶原引导骨组织再生膜及其结构、性能与细胞学[J]. 材料导报,2010,24(S1):385−388. [WU G L, REN T B, CAO C H, et al. Preparation of PLCL/gelatin and PLCL/collagen-guided bone tissue regeneration membranes by electrospinning method, as well as their structure, properties and cytology[J]. Materials Reports,2010,24(S1):385−388.

    WU G L, REN T B, CAO C H, et al. Preparation of PLCL/gelatin and PLCL/collagen-guided bone tissue regeneration membranes by electrospinning method, as well as their structure, properties and cytology[J]. Materials Reports, 2010, 24(S1): 385-388.
    [39]
    王佳冕. 静电纺林蛙皮胶原蛋白/PLLA纳米纤维膜及性能研究[D]. 长春: 吉林大学, 2015.

    WANG J M. Study on collagen/PLLA nanofiber membrane and its properties in electrospun forest frog skin[D]. Changchun: Jilin University, 2015.
    [40]
    肖世维, 但年华, 马文杰, 等. 胶原-聚乙烯醇共混纤维的电纺工艺条件探索[J]. 材料导报,2015,29(6):49−53,75. [XIAO S W, DAN N H, MA W J, et al. Exploration of electrospinning process conditions of collagen-polyvinyl alcohol blended fibers[J]. Materials Reports,2015,29(6):49−53,75.

    XIAO S W, DAN N H, MA W J, et al. Exploration of electrospinning process conditions of collagen-polyvinyl alcohol blended fibers[J]. Materials Reports, 2015, 29(6): 49-53, 75.
    [41]
    陈腊梅, 曹婕, 叶霖, 等. 同轴静电纺丝法制备具有核-壳纤维结构的多功能敷料[J]. 高等学校化学学报,2016,37(3):600−606. [CHEN L M, CAO J, YE LL, et al. Preparation of multifunctional dressings with core-shell fiber structure by coaxial electrospinning[J]. Chemical Journal of Chinese Universities,2016,37(3):600−606.

    CHEN L M, CAO J, YE LL, et al. Preparation of multifunctional dressings with core-shell fiber structure by coaxial electrospinning [J]. Chemical Journal of Chinese Universities, 2016, 37(3): 600-606.
    [42]
    陈卫英, 陈真勇, 杨在君, 等. 胶原-乙酸混合溶液静电纺丝可纺性及电纺胶原膜力学特性评估[J]. 材料导报,2021,35(S2):516−519. [CHEN W Y, CHEN Z Y, YANG Z J, et al. Evaluation of electrospinning spinnability and mechanical properties of electrospun collagen film of collagen-acetic acid mixed solution[J]. Materials Reports,2021,35(S2):516−519.

    CHEN W Y, CHEN Z Y, YANG Z J, et al. Evaluation of electrospinning spinnability and mechanical properties of electrospun collagen film of collagen-acetic acid mixed solution[J]. Materials Reports, 2021, 35(S2): 516-519.
    [43]
    雒永超. 胶原基纳米复合功能材料的制备及生物学评价研究[D]. 天津: 河北工业大学, 2018.

    LUO Y C. Preparation and biological evaluation of collagen-based nanocomposite functional materials[D]. Tianjin: Hebei University of Technology, 2018.
    [44]
    莫志超. 聚乳酸血管支架的载药及其生物相容性评价[D]. 青岛: 青岛科技大学, 2016.

    MO Z C. Drug loading and biocompatibility evaluation of polylactic acid vascular stents[D]. Qingdao: Qingdao University of Science and Technology, 2016.
  • Cited by

    Periodical cited type(3)

    1. 胡张捷,张宝贯,张智武. 固态胶原基材料在医疗器械中的应用. 中国组织工程研究. 2025(16): 3503-3512 .
    2. 姚丹丹,宋秘钊,韩洪帅,李家鑫,王彦珍. 胶原蛋白及其纳米纤维的制备与性能表征. 上海纺织科技. 2024(05): 38-45 .
    3. 孔伟帅,孙颖,王佳怡,李端鑫. 大麻纤维素的提取及其纳米纤维的制备与表征. 毛纺科技. 2024(12): 9-15 .

    Other cited types(0)

Catalog

    Article Metrics

    Article views (142) PDF downloads (26) Cited by(3)

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return