**4. Conclusions and future work**

**Figure 12.** Alcian blue staining for osteoinductive scaffolds. A different trend was observed in terms of acidic

**Figure 11.** Picrosirius red staining. Collagen synthesis was consistent and steady across all study groups.

140 Current Topics in the Utilization of Clay in Industrial and Medical Applications

According to the hypothesis, scaffold Groups 3 and 4 would induce differentiation of BSCs towards osteoblastic lineage as they contained different concentrations of DEX- doped HNTs. Hence, more alkaline phosphatase activity was predicted in these groups. Staining results of

mucopolysaccharide synthesis for all four groups.

*3.8.3. Alkaline phosphatase activity*

We conclude that CPCs fabricated using TTCP and DCPA as the solid phase and 10% chitosan lactate solution as the setting liquid and doped with HNTs have an osteoconductive nature as evidenced by a production of an abundant ECM synthesis and alkaline phosphatase activity. Surface roughness was increased with HNT addition which favors cell attachment, proliferation, and ECM synthesis. Halloysite increased the compressive strength of CPCs. The composition of CPCs after setting was analyzed using FTIR which reveals that the addition of HNTs increased HA production by acting as seeding agents for the precipitation of HA. CPC with DEX-doped HNTs showed an extended release of DEX from CPC scaffolds. DEX-loaded HNTs were used in varying concentrations in the fabrication of osteoinductive scaffolds. Osteoinductive scaffolds were seeded with MSCs to evaluate their osteoinductive nature. Our results suggest that HNTs with DEX loaded in the scaffolds started inducing differentiation in MSCs after 14 days while scaffolds with empty HNTs started inducing differentiation towards osteoblastic lineage after 3 days. At the end of 14th day, scaffolds containing non-loaded HNTs had the highest amount of alkaline phosphatase activity suggesting maximum differentiation of BSCs to osteoblastic phenotype. HNTs promoting differentiation of BSCs to osteoblastic phenotype can be explored further using different compositions. HNTs loaded with different drugs, antibiotics, growth factors, and chemotherapeutic agents can be used in fabricating CPCs. Addition of other polymers can also be considered to enhance the CPC mechanical properties further. *In vivo* experiments are being conducted in small animals to evaluate the tissue formation properties effectiveness of the CPCs fabricated.

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