To obtain a material with potential for use in tissue engineering, anionic collagen was obtained from porcine serosa (S) and bovine tendon (T) by alkaline hydrolysis for 72h. Part of this collagen was mixed with water to obtain 4 % (weight/weight) collagen suspension and part was solubilized in acetic acid pH 3.5 to obtain 1.5% (w/w) gel. The suspensions were mixed with their respective gels (2:1) (suspension: gel) and grape seed extract, whose main product is proanthocyanidin, was added at concentrations of 0.03% and 0.5%, thus obtaining the scaffolds SC (serosa collagen suspension and gel), TC (tendon collagen suspension and gel), SCP003 (SC with 0.03% extract), TCP003 (TC with 0.03% extract), SCP05 (SC with 0.5% extract added) and TCP05 (TC with 0.5% extract). The materials were analyzed by differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and characterized by phosphate buffered saline absorption assay and in vitro biological stability assay. By DSC it is observed that the addition of 0.5% of extract increases the denaturation temperature (Td) of collagen, indicating that at this concentration the extract acts as polymer crosslinking agent. SEM shows disorganized cross-section pores in all scaffolds, not exceeding 130 ?m. Absorption and degradation assays indicated that the addition of 0.5% extract increases the absorption of phosphate buffered saline (PBS) by the scaffolds and decreases the degradation percentage by collagenase. These results suggests that the scaffolds can be used for different applications, e.g. as hemostatic agent.
Tomizawa Y. Clinical benefits and risk analysis of topical hemostats: a review. J of Artificial Organs 8(3) : 137-142 (2005).
Carvalho M V H, Marchi E, Pantoroto M, Rossini M, Silva DMS, Teodoro LFF, Pantaroto A. Agentes hemostáticos locais e adesivos teciduais. Revista do Colégio Brasileiro de Cirurgiões 40(1) : 66-71 (2013).
Barnard J, Millner R. A Review of Topical Hemostatic Agents for Use in Cardiac Surgery. Annals of Thoracic. Surgery 88 : 1377–1383 (2009).
DeFrates KG, Moore R, Borgesi J, Lin G, Mulderig T, Beachley V, Hu X, Protein-Based Fiber Materials in Medicine: A Review. Nanomaterials 8 457 (2018).
Hess JR, Brohi K, Dutton RP, Hauser CJ, Holcomb JB, Kluger Y, The coagulopathy of trauma: a review of mechanisms. J Trauma 65 : 748–754 (2008).
Sundaram CP, Keenan AC Evolution of hemostatic agents in surgical practice. Indian J Urol. 26(3) : 374-78 (2010).
Junqueira LC, Carneiro J, Histologia Básica. ed 12. – Rio de Janeiro: Guanabara Koogan (2013).
Willett T L, Dapaah DY, Uppuganti S, Granke M, Nymanb JS. Bone collagen network integrity and transverse fracture toughness of human cortical bone. Bone, 120 : 187–193 (2019).
Sun J, Mou C, Shi Q, Chen B, Hou X, Zhang W, Li X, Zhuang Y, Shi J, Chen Y, Dai J. Controlled release of collagen-binding SFF-1α from collagen scaffold promoted tendon regeneration in a rat Achilles tendon defect model. Biomaterials, 162 : 22-33 (2018).
Bet MR, Goissis G, Lacerda CA. Characterization of polyanionic collagen prepared by selective hydrolysis of asparagine and glutamine carboxyamide side chains. Biomacromolecules, 2(4) : 1074-1079 (2001).
Goissis G, Suzigan S, Parreira DR, Maniglia JV, Braile DM, Raymundo SRO. Preparation and characterization of collagen-elastin matrices from blood vessels intended as small diameter vascular grafts. Artificial Organs, 24(3) : 217-223 (2000).
Rocha LB, Goissis G, Ross MA. Biocompatibility of Anionic Collagen Matrix as Scaffold for Boe Healing. Biomaterials, 23 : 449-456 (2002).
Forti FL, Goissis G, Plepis AMG. Modifications on Collagen Structures Promoted by 1,4-Dioxane Improve Thermal and Biological Properties of Bovine Pericardium as a Biomaterial. J of Biomaterials Applications, 20 : 267-285 (2006).
Chen Q, Liang S, Thouas GA. Elastomeric biomaterials for tissue engineering. Progress in Polymer Science, 38 : 584 – 671 (2013).
Han B, Jaurequi J, Tang BW, Nimni ME. Proanthocyanidin: A natural crosslinking reagent for stabilizing collagen matrices. J Biomedical Materials, 65 : 118-124 (2003).
Cho ML, Heo YJ, Park MK, Oh HJ, Park JS, Woo YJ, Ju JH. Grape seed proanthocyanidin extract (GSPE) attenuates collagen-induced arthritis. Immunology Letters, 124(2) : 102-110 (2009).
Horn MM, Martins VCA, Plepis AMG. Interaction of anionic collagen with chitosan: Effect on thermal and morphological characteristics. Carbohydrate Polymers, 77(2) : 239–243 (2009).
Allen, T. Particle Size Measurement- Powder sampling and particle size measurement. 5 ed, Londres, Chapman & Hall, pp 525 (1997).
Parenteau-Bareil R, Gauvin R, Berthod F. Collagen-Based Biomaterials for Tissue Engineering Applications. Materials, 3(3) : 1863-1887 (2010).
Lacerda C, Plepis AMG, Goissis G. Hidrólise seletiva de carboxiamidas de resíduos de asparagina e glutamina em colágeno: preparação e caracterização de matrizes aniônicas para uso como biomateriais, Quím Nova, 21(3) : 267-271 (1998).
Vidal CMP, Zhu W, Manohar S, Aydin B, Keiderling TA, Messersmith PB, Bedran-Russo AK. Collagen-collagen interactions mediated by plant-derived proanthocyanidins: A spectroscopic and atomic force microscopy study. Acta Biomaterialia, 41 : 110-118 (2016).
Nyberg E, Rindone A, Dorafshar A, Grayson WL. Comparison of 3D-Printed Poly-e-Caprolactone Scaffolds Functionalized with Tricalcium Phosphate, Hydroxyapatite, Bio-Oss, or Decellularized Bone Matrix. Tissue Engineering, 23 : 503-514 (2017).
Zhang K, Fan Y, Dunne N, Li X. Effect of microporosity on scaffolds for bone tissue enginnering. Regenerative Biomaterials 5 : 115-124 (2018).
Sionkowska A, Kaczmarek B, Lewandowska K. Modification of collagen and chitosan mixtures by the addition of tannic acid. J of Molecular Liquids, 199 : 318-323 (2014).
Walters BD, Stegemann JP. Strategies for directing the structure and function of three-dimensional collagen biomaterials across length scales. Acta Biomaterialia, 10 : 1488-1501 (2014).
Metzmacher I. Enzymatic degradation and drug release behavior of dense collagen implants. Dissertation – Department of Pharmacy, Pharmacetical Technology and Biopharmaceutics, Ludwig-Maximilians, University Munique, 2005.
Ma L, Gao C, Mao Z, Zhou J, Shen J, Hu X, Han C. Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering. Biomaterials, 24 : 4833-4841 (2003).
Rodrigues FT, Martins VCA, Plepis AMG. Porcine skin as a sources of biodegradable matrices: Alkaline treatment and glutaraldehyde crosslinking. Polímeros, 20(2) : 92-97 (2010).
Takigawa T, Endo Y. Effects of glutaraldehyde exposure on human health. J of Occupational Health, 48 : 75-87. (2006).