Consequences of incorporating thiaproline and its oxidized derivatives into collagen triple helices.

(2R)-4-thiaproline (Thp) is an analog of proline, replacing Cγ in the pyrrolidine ring with sulfur. Its thiazolidine ring easily interconverts between endo and exo puckers due to a small energy barrier, which leads to destabilize polyproline helices. Collagen, composed of three polyproline II helices, mainly consists of X-Y-Gly triplets, where X is often proline and Y is frequently (2S,4R)-hydroxyproline. In this study, we incorporated Thp into either position-X or position-Y to investigate the consequences of such a replacement on the triple helix. Circular dichroism and differential scanning calorimetry analyses showed that the Thp-containing collagen-mimetic peptides (CMPs) can fold into stable triple helices, in which the substitution at position-Y exhibits a larger destabilization effect. Additionally, we also prepared the derivative peptides by oxidizing Thp in the peptide to N-formyl-cysteine or S,S-dioxide Thp. The results showed that the oxidized derivatives at position-X only slightly affect collagen stability, but those at position-Y induce a large destabilization effect. The consequences of incorporating Thp and its oxidized derivatives into CMPs are position dependent. Computational results suggested that the ease of interconversion between exo and endo puckers for Thp and the twist conformation of S,S-dioxide Thp may cause the destabilization effect at position-Y. We have revealed new insights into the impacts of Thp and its oxidized derivatives on collagen and demonstrated that Thp can be used to design collagen-related biomaterials.

Peptoid Residues Make Diverse, Hyperstable Collagen Triple-Helices.

As the only ribosomally encoded N-substituted amino acid, proline promotes distinct secondary protein structures. The high proline content in collagen, the most abundant protein in the human body, is crucial to forming its hallmark structure: the triple-helix. For over five decades, proline has been considered compulsory for synthetic designs aimed at recapitulating collagen's structure and properties. Here we describe that N-substituted glycines (N-glys), also known as peptoid residues, exhibit a general triple-helical propensity similar to or greater than proline, enabling synthesis of stable triple-helical collagen mimetic peptides (CMPs) with unprecedented side chain diversity. Supported by atomic-resolution crystal structures as well as circular dichroism and computational characterizations spanning over 30 N-gly-containing CMPs, we discovered that N-glys stabilize the triple-helix primarily by sterically preorganizing individual chains into the polyproline-II helix. We demonstrated that N-glys with exotic side chains including a "click"-able alkyne and a photosensitive side chain enable CMPs for functional applications including the spatiotemporal control of cell adhesion and migration. The structural principles uncovered in this study open up opportunities for a new generation of collagen-mimetic therapeutics and materials.

A double-network polysaccharide-based composite hydrogel for skin wound healing.

Effective wound dressings are of great significance in preventing infections and promoting wound healing. However, most existing hydrogel dressings have an inadequacy in either mechanical performance, biological activities, or versatilities. Here we presented a double-network cross-linked polysaccharide-based hydrogel composed of collagen peptide-functionalized carboxymethyl chitosan (CS) and oxidized methacrylate sodium alginate (SA). The hydrogel possessed interconnected porous morphologies, suitable swelling ratios, excellent mechanical properties, and favorable biocompatibility. Meanwhile, the in vivo studies using a mouse full-thickness skin defect model showed that the double-network CS/SA hydrogel significantly accelerated wound healing by regulating the inflammatory process, promoting collagen deposition, and improving vascularization. Therefore, the functionalized double-network hydrogel should be a potential candidate as wound dressings.

Insertion of Pro-Hyp-Gly provides 2 kcal mol-1 stability but attenuates the specific assembly of ABC heterotrimeric collagen triple helices.

Collagen is a major structural component of the extracellular matrix and connective tissue. The key structural feature of collagen is the collagen triple helix, with a Xaa-Yaa-Gly (glycine) repeating pattern. The most frequently occurring triplet is Pro (proline)-Hyp (hydroxyproline)-Gly. The reversible thermal folding and unfolding of a series of heterotrimeric collagen triple helices with varying number of Pro-Hyp-Gly triplets were monitored by circular dichroism spectroscopy to determine the unfolding thermodynamic parameters Tm (midpoint transition temperature), ΔHTm (unfolding enthalpy), and ΔGunfold (unfolding free energy). The Tm and ΔGunfold of the heterotrimeric collagen triple helices increased with increasing number of Pro-Hyp-Gly triplets. The ΔGunfold increased by 2.0 ± 0.2 kcal mol-1 upon inserting one Pro-Hyp-Gly triplet into all three chains. The Tm difference between the most stable ABC combination and the second most stable BCC combination decreased with increasing number of Pro-Hyp-Gly triplets, even though the ΔGunfold difference remained the same. These results should be useful for tuning the stability of collagen triple helical peptides for hydrogel formation, recognition of denatured collagen triple helices as diagnostics and therapeutics, and targeted drug delivery.

Collagen-Based Thiol-Norbornene Photoclick Bio-Ink with Excellent Bioactivity and Printability.

As the essential foundation of bioprinting technology, cell-laden bio-ink is confronted with the inevitable contradiction between printability and bioactivity. For example, type I collagen has been widely applied for its excellent biocompatibility; however, its relatively low self-assembly speed restricts the performance in high-precision bioprinting of cell-laden structures. In this study, we synthesize norbornene-functionalized neutral soluble collagen (NorCol) by the reaction of acid-soluble collagen (Col) and carbic anhydride in the aqueous phase. NorCol retains collagen triple-helical conformation and can be quickly orthogonally cross-linked to build a cell-laden hydrogel via a cell-friendly thiol-ene photoclick reaction. Moreover, the additional carboxyl groups produced in the reaction of carbic anhydride and collagen obviously improve the solubility of NorCol in neutral buffer and miscibility of NorCol with other polymers such as alginate and gelatin. It enables hybrid bio-ink to respond to multiple stimuli, resulting in continuous cross-linked NorCol networks in hybrid hydrogels. For the first time, the collagen with a triple helix structure and gelatin can be mixed and printed, keeping the integrity of the printed construct after gelatin's dissolution. The molecular interaction among giant collagen molecules allows NorCol hydrogel formation at a low concentration, which leads to excellent cell spreading, migration, and proliferation. These properties give NorCol flexible formability and excellent biocompatibility in temperature-, ion-, and photo-based bioprinting. We speculate that NorCol is a promising bio-ink for emerging demands in tissue engineering, regenerative medicine, and personalized therapeutics.

Feasibility Study of Gelatin Preparation from the Bioinspired Collagen Aggregates by a "Two-step" Facile Degradation Method.

Gelatin is the putative research hotspot of natural products, but gelatin prepared by traditional alkali methods has seriously affected its applications due to the worryingly low molecular weight and poor gel strength. Herein, we took the lead to extract the distinct gelatin from a kind of bioinspired collagen aggregate (CA) by a two-step controlled degradation method. Structural analysis suggested that the CA better preserves the natural aggregated structure of nature collagen (typical D-periodic cross-striated pattern). Compared with the gelatin gelatinized by the conventional alkali method (G-Al) and commercial gelatin (CG), the gelatin (G-CA) from CA had a wide molecular weight distribution range, high transparency, high viscosity, and strong gel strength as expected. Meanwhile, the G-CA film exhibited better mechanical performance and thermostability than CG and G-Al films, and water vapor permeability was also higher in the G-CA film, whereas water solubility was higher in the CG and G-Al films. Thus, the G-CA film is more conducive to the use of food packaging or edible films, exhibiting more potential market application prospects. Notably, G-CA based on CA from waste hide offal provides a way to reuse leather waste resources and further realize green and clean production in leather industry.

Surface-Directed Mineralization of Fibrous Collagen Scaffolds in Simulated Body Fluid for Tissue Engineering Applications.

The use of polymer additives that stabilize fluidic amorphous calcium phosphate is key to obtaining intrafibrillar mineralization of collagen in vitro. On the other hand, this biomimetic approach inhibits the nucleation of mineral crystals in unconfined extrafibrillar spaces, that is, extrafibrillar mineralization. The extrafibrillar mineral content is a significant feature to replicate from hard connective tissues such as bone and dentin as it contributes to the final microarchitecture and mechanical stiffness of the biomineral composite. Herein, we report a straightforward route to produce densely mineralized collagenous composites via a surface-directed process devoid of the aid of polymer additives. Simulated body fluid (1×) is employed as a biomimetic crystallizing medium, following a preloading procedure on the collagen surface to quickly generate the amorphous precursor species required to initiate matrix mineralization. This approach consistently leads to the formation of extrafibrillar bioactive minerals in bulk collagen scaffolds, which may offer an advantage in the production of osteoconductive collagen-apatite materials for tissue engineering and repair purposes.

Design of hydrogel-based scaffolds for the treatment of spinal cord injuries.

Spinal cord injury (SCI) is a traumatic lesion that diminishes sensory and/or motor neuronal functionality, directly affecting the quality of the patient's life. Due to the central nervous system's (CNS) inhibitory microenvironment that presents challenges in neuron repair and regeneration, tissue engineering strategies have received significant attention to improve the quality of a patient's life. In this regard, hydrogels are attractive SC scaffolds as they can provide not only an adjustable physiologically native-like microenvironment but also an appropriate matrix for cell delivery, drug delivery, and other bioactive molecule delivery at the lesion site. This systematic review characterizes the widely used biomaterials including natural polymers; protein- and polysaccharide-based synthetic polymers; methacrylate- and polyethylene glycol-based, and self-assembling (SA) peptides. In addition, synthesis routes of hydrogels are investigated. This review is complemented by the discussion of the various techniques utilized for hydrogel scaffold designs with their in vitro and in vivo outcomes and clinical trials. The existing challenges and opportunities for SC hydrogel scaffolds are mentioned towards the end of this review.

A graphene oxide-aided triple helical aggregation-induced emission biosensor for highly specific detection of charged collagen peptides.

Aggregation-induced emission (AIE) probes have emerged as promising "turn-on" sensing tools for DNA and proteins, and the AIE biosensors conjugated with graphene oxide (GO) have shown improved selectivity. Collagen is an essential structural protein in the human body, and its degraded products are involved in a plethora of severe diseases. Collagen has a high content of charged amino acids, while EOG represents one of the most abundant charged triplets in Type I collagen. We, herein, for the first time report the construction of a GO-aided AIE biosensor for the detection of charged collagen peptides. We have shown that an AIE fluorophore TPE conjugated with a triple helical peptide TPE-PRG possesses strong fluorescence due to the restriction of intramolecular rotation of TPE in the trimer state. The adsorption of the probe TPE-PRG by GO leads to efficient fluorescence quenching, while the addition of target collagen peptide EOG releases the probe peptide from the GO surface and recovers its fluorescence. We have demonstrated that the TPE-PRG/GO complex provides a highly specific "turn-on" sensing platform for the target collagen peptide with a typical charged amino acid-rich sequence. The assay has shown little interference from other biomolecules, and it can also effectively distinguish the target charged collagen peptide from its single amino acid mutant type. The development of robust analytical assays for charged collagen peptides could pronouncedly extend our capability to investigate the pathology of collagen diseases, showing great potential for their molecular diagnosis.

Glutamic acid concentration dependent collagen mineralization in aqueous solution.

The aim of this research is to evaluate the effects of glutamic acid (Glu) on the process of collagen mineralization, the structure and property of mineralized composites. Collagen mineralization was initiated by introducing PBS to blended solutions containing 1 mg/mL collagen, 6 mmol/L calcium and Glu ranged from 0 to 550 mmol/L. The kinetic curves and quantitation analyses showed that Glu could delay the collagen mineralization, and reduce the crystalline size and the amount of hydroxyapatite. With the Glu concentration increased from 50 to 200 mmol/L, the collagen self-assembly was promoted, resulting in the improvement of hardness and thermal stability of mineralized composites. However, further increase in the Glu concentration to 400 mmol/L or above would significantly inhibit the self-assembly of collagen and reduce the hardness and thermal stability of mineralized composites. Scanning electron microscopy revealed that the diameter of collagen became thicker as the Glu concentration increased. Moreover, hydroxyapatite with spherical morphology was uniformly dispersed and well combined with collagen fibril at Glu concentration of 200 mmol/L. These results may provide a broader understanding of the potential mechanism of biomineralization and be critical in the design of biomimetic scaffolds for bone tissue engineering.

Zinc Silicate/Nano-Hydroxyapatite/Collagen Scaffolds Promote Angiogenesis and Bone Regeneration via the p38 MAPK Pathway in Activated Monocytes.

Recent studies show that biomaterials are capable of regulating immune responses to induce a favorable osteogenic microenvironment and promote osteogenesis and angiogenesis. In this study, we investigated the effects of zinc silicate/nanohydroxyapatite/collagen (ZS/HA/Col) scaffolds on bone regeneration and angiogenesis and explored the related mechanism. We demonstrate that 10ZS/HA/Col scaffolds significantly enhanced bone regeneration and angiogenesis in vivo compared with HA/Col scaffolds. ZS/HA/Col scaffolds increased tartrate-resistant acid phosphatase (TRAP)-positive cells, nestin-positive bone marrow stromal cells (BMSCs) and CD31-positive neovessels, and expression of osteogenesis (Bmp-2 and Osterix) and angiogenesis-related (Vegf-α and Cd31) genes increased in nascent bone. ZS/HA/Col scaffolds with 10 wt % ZS activated the p38 signaling pathway in monocytes. The monocytes subsequently differentiated into TRAP+ cells and expressed higher levels of the cytokines SDF-1, TGF-ß1, VEGF-α, and PDGF-BB, which recruited BMSCs and endothelial cells (ECs) to the defect areas. Blocking the p38 pathway in monocytes reduced TRAP+ differentiation and cytokine secretion and resulted in a decrease in BMSC and EC homing and angiogenesis. Overall, these findings demonstrate that 10ZS/HA/Col scaffolds modulate monocytes and, thereby, create a favorable osteogenic microenvironment that promotes BMSC migration and differentiation and vessel formation by activating the p38 signaling pathway.

Biomimetic mineralizable collagen hydrogels for dynamic bone matrix formation to promote osteogenesis.

The simulation of the native bone matrix formation process is crucial for the construction of the cellular microenvironment for bone regeneration. However, it is still challenging to design bioactive materials that simultaneously mimic the composition and dynamic mineralization process of the bone matrix, let alone realize osteoinduction by a biomimetic dynamic microenvironment. In this study, we prepared a biomimetic mineralizable collagen hydrogel (CAV) and explored the effects of a dynamic mineralized matrix on the osteogenesis of stem cells both in vitro and in vivo. We showed the feasibility of the biomimetic CAV hydrogel to induce mineralization in simulated media including simulated body fluid (SBF), glycerol phosphate calcium salt hydrate (CaGP) solution and cell co-cultured systems. The participation of cells in the mineralization process is more likely to induce matrix remodeling due to the synergistic effects of CAV mineralization and cellular secretion, resulting in higher matrix strength. We also demonstrated that the biomimetic mineralized hydrogel could up-regulate osteogenic genes and protein expression of bone marrow mesenchymal stem cells (BMSCs), thus enhancing osteogenesis in vivo. The interactions between the mineralizable hydrogel and cells play an important role in regulating dynamic matrix mineralization and osteogenesis. Our findings prove that the biomimetic mineralizable hydrogel is a promising candidate for implantable orthopedic applications and provides essential implications for the future design of materials for bone regeneration.

Newly Designed Human-Like Collagen to Maximize Sensitive Release of BMP-2 for Remarkable Repairing of Bone Defects.

Designing the "ideal" hydrogel/matrix which can load bone morphogenetic protein-2 (BMP-2) in a low dose and with a sustained release is the key for its successful therapeutic application to enhance osteogenesis. The current use of natural collagen sponges as hydrogel/matrix is limited due to the collagen matrix showing weak mechanical strength and unmanageable biodegradability. Furthermore, the efficiency and safe dose usage of the BMP-2 has never been seriously considered other than purely chasing the lowest dose usage and extended-release time. In this paper, we customized a novel enzymatically cross-linked recombinant human-like collagen (HLC) sponge with low immunogenicity, little risk from hidden viruses, and easy production. We obtained a unique vertical pore structure and the porosity of the HLC, which are beneficial for Mesenchymal stem cells (MSCs) migration into the HLC sponge and angiopoiesis. This HLC sponge loading with low dose BMP-2 (1 µg) possessed high mechanical strength along with a burst and a sustained release profile. These merits overcome previous limitations of HLC in bone repair and are safer and more sensitive than commercial collagens. For the first time, we identified that a 5 µg dose of BMP-2 can bring about the side effect of bone overgrowth through this sensitive delivery system. Osteoinduction of the HLC-BMP sponges was proved by an in vivo mouse ectopic bone model and a rat cranial defect repair model. The method and the HLC-BMP sponge have the potential to release other growth factors and aid other tissue regeneration. Additionally, the ability to mass-produce HLC in our study overcomes the current supply shortage, which limits bone repair in the clinic.

Sustained release of ketorolac tromethamine from poloxamer 407/cellulose nanofibrils graft nanocollagen based ophthalmic formulations.

There has been extensive utilization of poloxamer 407 (PM) for the delivery of various ophthalmic drugs aimed at efficient ophthalmic drug delivery approach for longer precorneal residence time along with acceptable bioavailability of drugs. We have studied the effect of nanocellulose grafted collagen (CGC) on the performance of in situ gels based on PM for the controlled in vitro release of Ketorolac Tromethamine (KT). CGC has shown great influence evident by the reduction in PM critical gelation concentration, increased gel strength, and prolonged the release of loaded drugs compared with the virgin PM gel. The engineered nanocomposite formulations established an anomalous diffusion mechanism along with a Fickian diffusion controlled drug release for 1.5 & 1.75 w/v% CGC reinforced PM. Hence, the synthesized in situ nanocomposites are potential candidates for ophthalmic drug delivery system.

Preparation and characterization of the collagen/cellulose nanocrystals/USPIO scaffolds loaded kartogenin for cartilage regeneration.

The regeneration of cartilage is a challenging problem for lack of innate abilities to mount a sufficient healing response. Kartogenin (KGN), an emerging chondroinductive non-protein small molecule, bound to the surface of the ultrasmall super-paramagnetic iron-oxide (USPIO) by innovational one-step technology, followed by being incorporated into the cross-linking collagen/cellulose nanocrystals (Col/CNC) bioactive scaffolds to stimulate an appropriate microenvironment for the growth and differentiation of bone marrow-derived mesenchymal stem cells (BMSCs), thus facilitating the formation of chondrocyte. Herein, USPIO not only served as a carrier for small molecule drugs, but also as MRI contrast agents, which can non-invasively monitor the degradation of the scaffolds and the self-repair capacity of cartilage. In vitro studies showed that the KGN could release from the composite scaffolds in a sustained and stable manner and promote the chondrogenic differentiation of BMSCs based on UV spectrophotometry test, and specific markers analysis. Of note, USPIO labeled composite scaffolds retained their stability without loss of relaxation rate the composite scaffolds can be a promising biomaterials for cartilage repair, with the function of noninvasive visualization and semiquantitative analysis of scaffolds degradation and neocartilage.

Modification of chitosan grafted with collagen peptide by enzyme crosslinking.

Free radicals are closely related to the occurrence and development of aging, cancer and inflammation. In this paper, the microbial transglutaminase (MTGase) was used as a catalyst to graft the collagen peptide (COP) molecules on the amino group of carboxymethyl chitosan sulfate (CMCS) to improve the antioxidant effects. FT-IR and NMR spectroscopy were used to confirm the successful grafting of COP to CMCS. Degree of substitution (DS) of CMCS-COP could be controlled by adjusting the reaction conditions. With the increase of concentration, the ability of each sample on scavenging capacity and reducibility tends to increase obviously. The results of anticoagulant experiments showed that the ability of CMCS and CMCS-COP with three different degrees of substitution on activated partial thrombin time (APTT) and prothrombin time (PT) values were all increased to compare with the control group. No relevant cytotoxicity against NIH-3T3 mouse fibroblasts was found for the copolymers. These results suggested that CMCS-COP would appear to be a promising candidate for wound dressing application.

Preparation and in vitro characterization of cross-linked collagen-gelatin hydrogel using EDC/NHS for corneal tissue engineering applications.

Corneal disease is considered as the second leading cause of vision loss and keratoplasty is known as an effective treatment for it. However, the tissue engineered corneal substitutes are promising tools in experimental in vivo repair of cornea. Selecting appropriate cell sources and scaffolds are two important concerns in corneal tissue engineering. The object of this study was to investigate biocompatibility and physical properties of the bio-engineered cornea, fabricated from type-I collagen (COL) and gelatin (Gel). Two gelatin based hydrogels cross-linked with EDC/NHS were fabricated, and their physicochemical properties such as equilibrium water content, enzymatic degradation, mechanical properties, rheological, contact angle and optical properties as well as their ability to support human bone-marrow mesenchymal stem cells (hBM-MSCs) survival were characterized. The equilibrium water content and enzymatic degradation of these hydrogels can be easily controlled by adding COL. Our findings suggest that incorporation of COL-I increases optical properties, hydrophilicity, stiffness and Young's modulus. The viability of hBM-MSCs cultured in Gel and Gel: COL was assessed via CCK-8 assay. Also, the morphology of the hBM-MSCs on the top of Gel and Gel: COL hydrogels were characterized by phase-contrast microscopy. This biocompatible hydrogel may promise to be used as artificial corneal substitutes.

Preparation and characterization of novel poly (vinyl alcohol)/collagen double-network hydrogels.

Hydrogels prepared by conventional methods show poor performance in many aspects. Many double network (DN) hydrogels prepared by crosslinking agents have disadvantages such as toxicity. In this work, we prepared novel DN hydrogels with fish-originated collagen (Col) and poly (vinyl alcohol) (PVA), in which self-assembly of collagen and self-crosslinking of PVA were achieved. Infrared spectra indicated the existence of double network with chemical interactions between Col and PVA. X-ray diffraction (XRD) patterns showed that characteristic peak of freeze-thaw PVA in DN hydrogel was retained. Scanning electron microscope (SEM) images before and after degradation and swelling property tests indicated that the morphology of the hydrogel was a compact meshwork, which is consistent with the high water-retention rate. The degradation rate of the DN hydrogels was controlled by the ratio of Col and PVA. Compared with a pure collagen hydrogels, the stress of DN hydrogels was greatly enhanced from 6 to 33 kPa at a strain of 40%. This study indicates that DN hydrogels prepared by Col and PVA are an ideal biomaterial for tissue engineering.

Biomimetic hydrogels direct spinal progenitor cell differentiation and promote functional recovery after spinal cord injury.

OBJECTIVE: Demyelination that results from disease or traumatic injury, such as spinal cord injury (SCI), can have a devastating effect on neural function and recovery. Many researchers are examining treatments to minimize demyelination by improving oligodendrocyte availability in vivo. Transplantation of stem and oligodendrocyte progenitor cells is a promising option, however, trials are plagued by undirected differentiation. Here we introduce a biomaterial that has been optimized to direct the differentiation of neural progenitor cells (NPCs) toward oligodendrocytes as a cell delivery vehicle after SCI. APPROACH: A collagen-based hydrogel was modified to mimic the mechanical properties of the neonatal spinal cord, and components present in the developing extracellular matrix were included to provide appropriate chemical cues to the NPCs to direct their differentiation toward oligodendrocytes. The hydrogel with cells was then transplanted into a unilateral cervical contusion model of SCI to examine the functional recovery with this treatment. Six behavioral tests and histological assessment were performed to examine the in vivo response to this treatment. MAIN RESULTS: Our results demonstrate that we can achieve a significant increase in oligodendrocyte differentiation of NPCs compared to standard culture conditions using a three-component biomaterial composed of collagen, hyaluronic acid, and laminin that has mechanical properties matched to those of neonatal neural tissue. Additionally, SCI rats with hydrogel transplants, with and without NPCs, showed functional recovery. Animals transplanted with hydrogels with NPCs showed significantly increased functional recovery over six weeks compared to the media control group. SIGNIFICANCE: The three-component hydrogel presented here has the potential to provide cues to direct differentiation in vivo to encourage regeneration of the central nervous system.

Development of various composition multicomponent chitosan/fish collagen/glycerin 3D porous scaffolds: Effect on morphology, mechanical strength, biostability and cytocompatibility.

The various composition multicomponent chitosan/fish collagen/glycerin 3D porous scaffolds were developed and investigated the effect of various composition chitosan/fish collagen/glycerin on scaffolds morphology, mechanical strength, biostability and cytocompatibility. The scaffolds were fabricated via freeze-drying technique. The effects of various compositions consisting in 3D scaffolds were investigated via FT-IR analysis, porosity, swelling and mechanical tests, and effect on the morphology of scaffolds investigated microscopically. The biostability and cytocompatibility tests were used to explore the ability of scaffolds to use for tissue engineering application. The average pore sizes of scaffolds were in range of 100.73±27.62-116.01±52.06, porosity 71.72±3.46-91.17±2.42%, tensile modulus in dry environment 1.47±0.08-0.17±0.03MPa, tensile modulus in wet environment 0.32±0.03-0.14±0.04MPa and biodegradation rate (at day 30) 60.38±0.70-83.48±0.28%. In vitro culture of human fibroblasts and keratinocytes showed that the various composition multicomponent 3D scaffolds were good cytocompatibility however, the scaffolds contained high amount of fish collagen excellently facilitated cell proliferation and adhesion. It was found that the high amount fish collagen and glycerin scaffolds have high porosity, enough mechanical strength and biostability, and excellent cytocompatibility.