Poly(D,L-lactide acid, PDLLA) has been researched for scaffolds in bone regeneration. However, its hydrophobocity and smooth surface impedes its interaction with biological fluid and cell adhesion. To alter the surface characteristics, different surface modification techniques have been developed to facilitate biological application. The present study compared two different routes to produce PDLLA/superhydrophilic vertically aligned carbon nanotubes:nanohydroxyapatite (PDLLA/VACNT-O:nHAp) scaffolds. For this, we used electrodeposition and immersion in simulated body fluid (SBF). Characterization by goniometry, scanning electron microscopy, X-ray diffraction, and infrared spectroscopy confirmed the polymer modifications, the in vitro bioactivity, and biomineralization. Differential scanning calorimetry and thermal gravimetric analyses showed that the inclusion of VACNT-O:nHA probably acts as a nucleating agent increasing the crystallization rate in the neat PDLLA without structural alteration. Our results showed the formation of a dense nHAp layer on all scaffolds after 14 days of immersion in SBF solution; the most intense carbonated nHAp peaks observed in the PDLLA/VACNT-O:nHAp samples suggest higher calcium precipitation compared to the PDLLA control. Both cell viability and alkaline phosphatase assays showed favorable results, because no cytotoxic effects were present and all produced scaffolds were able to induce detectable mineralization. Bone defects were used to evaluate the bone regeneration; the confocal Raman and histological results confirmed high potential for bone applications. In vivo study showed that the PDLLA/VACNT-O:nHAp scaffolds mimicked the immature bone and induced bone remodeling. These findings indicate surface improvement and the applicability of this new nanobiomaterial for bone regenerative medicine.

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An association between in vitro and in vivo studies has been demonstrated for the first time, using a novel nanohydroxyapatite/superhydrophilic vertically aligned multiwalled carbon nanotube (nHAp/VAMWCNT-O2) nanocomposites. Human osteoblast cell culture and bone defects were used to evaluate the in vitro extracellular matrix (ECM) calcification process and bone regeneration, respectively. The in vitro ECM calcification process of nHAp/VAMWCNT-O2 nanocomposites were investigated using alkaline phosphatase assay. The in vivo biomineralization studies were carried out on bone defects of C57BL/6/JUnib mice. Scanning electron microscopy, micro-energy dispersive spectroscopy, X-ray photoelectron spectroscopy, and X-ray difractometry analyses confirmed the presence of the nHAp crystals. nHAp/VAMWCNT-O2 nanocomposites induced in vitro calcification of the ECM of human osteoblast cells in culture after only 24 h. Bone regeneration with lamellar bone formation after 9 weeks was found in the in vivo studies. Our findings make these new nanocomposites very attractive for application in bone tissue regeneration.

Polymer membranes have been widely used in guided tissue regeneration (GTR) and guided bone regeneration (GBR). The literature recognizes that poly (lactic acid) (PLA)/poly (ε-caprolactone) (PCL) blends have better physicochemical properties and that a porous polymer surface facilitates cell adhesion and proliferation. In addition, hydroxyapatite (HAp) incorporated into the polymer matrix promotes osteoinduction properties and osteoconduction to the polymer-ceramic biocomposite. Therefore, polymer membranes of PLA/PCL blend with the addition of HAp could be an alternative to be used in GBR. HAp was obtained by precipitation using the mixture of solutions of tetrahydrate calcium nitrate and monobasic ammonium phosphate salts. The porous membranes of the PLA/PCL (80/20) blend with the addition of HAp were obtained by solvent casting with a controlled humidity method, with the dispersion of HAp in chloroform and subsequent solubilization with the components of the blend. The solution was poured into molds for solvent evaporation under a controlled humidity atmosphere. The membranes showed the formation of pores on their surface, together with dispersed HAp particles. The results showed an increase in the surface porosity and improved bioactivity properties with the addition of HAp. Moreover, in biological studies with cell culture, it was possible to observe that the membranes with HAp have no cytotoxic effect on MC3T3 cells. These results indicate a promising use of the new biomaterial for GBR.

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