Table 1.
Primer sequences used for the detection of the genes.
Fig 1.
Surface Morphology (a) Spongy Scaffold (BCP) (b) 0.5% Alg-1% CHT-BCP (BCP-PEL Hy) (c) 0.5% Alg-1% CHT-BCP-BMP-2 (BCP-PEL Hy- BMP 2) (d) 12mm Scaffold for In Vivo (e) BCP loaded Chitosan at core (f) EDS profile for spongy scaffold.
Fig 2.
FTIR transmittance and XRD of the fabricated scaffolds.
Fig 3.
3D model for pore size orientation (a) Spongy Scaffold (BCP) (b) 0.5% Alg-1% CHT-BCP (BCP-PEL Hy) (c) 0.5% Alg-1% CHT-BCP-BMP-2 (BCP-PEL Hy-BMP2).
Table 2.
Physical characterization.
Fig 4.
(A) Lysozyme activated degradation Behavior of BCP-PEL Hydrogel system. (B) Degradation behavior.
Fig 5.
Cumulative BMP-2 release from BCP-PEL HY-BMP 2 system.
Fig 6.
Cell Viability and Proliferation of spongy Scaffold: (a) MTT Assay (*, P<0.05; ** P<0.01; *** P<0.001) (b) Live dead Assay (c) F-actin Assay.
Fig 7.
(A) The expression of osteopontin (OPN); alkaline phosphate (ALP); osteocalcin (OCN) and collagen type 1 (COL I) gene of MC3T3-E1cells for BCP; BCP-PEL hy and BCP-PEL hy-BMP 2 after 7 days and 14 days of cell culture in osteogenesis media, observed by real-time PCR. (*p< 0.05; **p<0.01; ***p<0.001; ns: no significant different). (B) The expression of osteopontin (OPN); alkaline phosphate (ALP) and collagen type 1 (COL I) protein of MC3T3-E1cells for BCP (1, 4); BCP-PEL hy (2, 5) and BCP-PEL hy-BMP 2 (3, 6) after 7 days and 14 days of cell culture in osteogenesis media, observed by Western Blot. β-actin levels are used as internal control.
Fig 8.
3D Model for new bone in growth from CT analysis for 4W and 8 W: Radiographic image and bone in growth of (a) BCP-PEL Hy-BMP 2 (b) BCP-PEL Hy (c) BCP; Quantitative data: (d) Bone volume/Tissue volume ratio (BV/TV) (e) Trabecular thickness (Tb. Th) (f) Trabecular number (Tb.N). (*p< 0.05; **p<0.01; ***p<0.001; ns: no significant different).
Fig 9.
(A)(a) Schematic diagram of implantation (b) Actual implantation Site; (B): New bone in growth by H&E staining for 4W and 8W. Here HB, NB, BM and ST stands for host bone, new bone, bone marrow and soft tissue. Encircled region is enlarged in Figs 10 and 11.
Fig 10.
Cellular activity at scaffold-host bone interphase at 4 W time period.
Here HB, NB, BM, CT, FB and COL stands for host bone, new bone, bone marrow, cartilage template and collagen.
Fig 11.
Cellular activity at scaffold-host bone interphase at 8 W time period.
Here HB, NB, BM, CT, FB and COL stands for host bone, new bone, bone marrow, cartilage template and collagen.