title: Biocomposite cryogels as tissue-engineered biomaterials for regeneration of critical-sized cranial bone defects.
abstract: Analysis of the in vivo regeneration capability of any tissue-engineered biomaterial is necessary once it shows potential characteristics during in vitro studies. Thus, we applied polyvinyl alcohol-tetraethylorthosilicate-alginate-calcium oxide (PTAC) biocomposite cryogel on critical-sized cranial bone defects in wistar rats for examining the comparative bone regeneration of cryogel-treated and nontreated defects over a period of 4 weeks. An in-depth analysis was performed from macroscopic level till the gene level. Bone regeneration in cryogel-treated defects was clearly evident from the results, whereas the nontreated group did not show any defect healing except at few peripheral areas. At the macroscopic level, micro-computed tomography analysis revealed new bone formation. This was further confirmed at the cellular level, wherein, new bone formation was demonstrated by hematoxylin and eosin staining. Osteoblastic differentiation was further validated by immunohistological staining of runt-related transcription factor-2 (Runx-2) protein and via calcium-phosphate crystal formation after 2 weeks through scanning electron microscopy and energy dispersive X-ray spectroscopy. Finally, at the gene level, real-time PCR analysis confirmed the mRNA expression of osteoblastic markers, that is, runx-2, collagen type I (Col I), alkaline phosphatase (ALP), and osteocalcin (OCN). Therefore, the results of in vivo cranial defect model studies suggest that PTAC biocomposite cryogels can show suitable potential for human bone regeneration.
authors.affiliation: 1 Department of Biological Sciences and Bioengineering, Indian Institute of Technology Kanpur , Kanpur, India .
authors.forename: Ruchi
authors.initials: R
authors.lastname: Mishra
authors.forename: Sudhir Kumar
authors.initials: SK
authors.lastname: Goel
authors.forename: Kailash Chand
authors.initials: KC
authors.lastname: Gupta
authors.forename: Ashok
authors.initials: A
authors.lastname: Kumar
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000494:pharmacology*; D001861:Bone Regeneration / Q000187:drug effects*; D002130:Calcium Phosphates / Q000494:pharmacology; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D059985:Cryogels / Q000494:pharmacology*; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D012886:Skull / Q000000981:diagnostic imaging / Q000187:drug effects / Q000473:pathology* / Q000503:physiopathology*; D012984:Software; D023822:Tissue Engineering / Q000379:methods*; D055114:X-Ray Microtomography
chemical_list: D001672:Biocompatible Materials; D002130:Calcium Phosphates; D050659:Core Binding Factor Alpha 1 Subunit; D059985:Cryogels; C020243:calcium phosphate