Characterization of human primary osteoblast response on bioactive glass (BaG 13-93)- coated poly-L,DL-lactide (SR-PLA70) surface in vitro.

Bioabsorbable polylactide-based polymers are commonly used for bone reconstruction. Although these polymers have proven successful in many applications, they do not have the capacity to induce osteoconduction. Therefore, several strategies have been developed to manufacture osteoconductive polylactide-based composites. In this study, we have investigated in vitro response of human primary osteoblasts for self-reinforced poly-L,DL-lactide 70/30 (SR-PLA70) plates coated with spheres of bioactive glass 13-93 (SR-PLA70 + BaG). Osteoblasts were cultured on SR-PLA70 and SR-PLA70 + BaG plates for 2, 7, or 14 days. By day 7, both materials induced a reduction in total cell population. However, by day 14 the proliferative response of osteoblasts on SR-PLA70 + BaG surface was such that the cell population had regained similar levels as that of day 2 controls. Alkaline phosphatase activity was higher on SR-PLA70 at day 7 but declined to control levels by day 14. There were no significant time-dependent variations in alkaline phosphatase activity on SR-PLA70 + BaG. After in vitro hydrolysis for 7 days, the elemental analysis of SR-PLA70 + BaG surface showed the presence of mineral precipitates that were confirmed as crystalline hydroxyapatite. This was accompanied by osteoblast spreading, protrusions of microvilli adhered to BaG 19-39 surface, cuboidal phenotype and cell surface associated formation of hydroxyapatite microspheres. In conclusion, the SR-PLA70 + BaG composite is capable of inducing a proliferative response of human primary osteoblasts, and appears to support the development of mature osteoblast phenotype. Therefore, the SR-PLA70 + BaG composites appear as promising osteoconductive scaffold candidates for reconstruction and regeneration of bone matrix.

Biodegradable polydioxanone and poly(l/d)lactide implants: an experimental study on peri-implant tissue response.

Several implants for orbital wall fracture treatment are available at the present, but they have drawbacks: resorption, risk for migration and foreign body reaction. Alloplastic resorbable implants would be advantageous: no removal operation and no donor side morbidity. The purpose of this study was to evaluate the foreign body reaction, capsule formation and mechanical properties of two bioresorbable implants. PDS and SR-P(L/DL)LA mesh sheet (70/30) with solid frame (96/4) implants (SR-P(L/DL)LA 70,96) were placed into subcutaneous tissue of 24 rats. Immunohistochemistry was used to evaluate reactivity for Tn-C, alpha-actin, type I and III collagens and two mononuclear cells: T-cells and monocyte/ macrophage. GPC, DSC and SEM were performed. Student's t-test or nonparametric Kruskall-Wallis test were used for statistical analysis. Histology of peri-implant capsule exhibited an inner cell-rich zone and an outer connective tissue zone around both materials. Tn-C reactivity was high in the inner and alpha-actin in the outer zone. At the end of the study, the difference of type I collagen versus type III collagen reactivity in inner zone was statistically significant (P<0.0001) as was the difference of type I collagen versus type III collagen reactivity in outer zone (P<0.0001). Immunohistochemistry did not reveal any statistical differences of T-cell and monocyte/macrophage reactivity around PDS versus SR-P(L/DL)LA 70,96 implants, nor any differences as a function of time. PDS were deformed totally after 2 months. SR-P(L/DL)LA 70,96 implants were only slightly deformed during the follow up of 7 months. PDS degraded rapidly in SEM observation. Particles were detaching from surface. SEM observation revealed that polylactide implant was degrading from the surface and the inner porous core became visible. The degradation came visible at 7 months. There were cracks in perpendicular direction towards to the long axis of the filaments. M(w) of PDS decreased fast compared to the polylactide implant. Foreign body reaction was minimal to both materials but continued throughout the whole observation period. Mechanically PDS was poor, it looses its shape totally within 2 months. It cannot be recommended for orbital wall reconstruction. New mesh sheet-frame structure (SR-P(L/DL)LA 70,96) approved to be mechanically adequate for orbital wall reconstruction. It seems not to possess intrinsic memory and retains its shape. The resorption time is significantly longer compared to PDS and is comparable to other studied P(L/DL)LA copolymers. Thus, the new polylactide copolymer implant may support the orbital contents long enough to give way to bone growth over the wall defect.

Vascular damage and lack of angiogenesis in systemic sclerosis skin.

The aim of this study was to analyse microvascular damage and compensatory angiogenesis in skin from patients with systemic sclerosis (SSc) compared with systemic lupus erythematosus (SLE), Raynaud's phenomenon (RP) and healthy controls. Immunohistochemistry was used for skin biopsies (9 SSc, 10 SLE, 9 RP and 12 healthy controls) using von Willebrand factor and beta3 integrin subunit specific antibodies, TechMate immunostaining robot and biotin-streptavidin protocol. In the early stages of SSc, vWF was found in the perivascular space and interstitial matrix in papillary but not in the reticular dermis, in particular around small oedematous blood vessels infiltrated by mononuclear cells. The extravascular release of vWF in SSc specimens was associated with weak or even a total lack of immunoreactivity within the associated endothelial cells. Late stages of SSc were characterised by loss of the dermal papillae, subepidermal fibrosis, hypovascularity and strong endothelial vWF expression without extravascular leakage. In all SSc patients studied only a few vascular profiles were weakly immunostained for beta3 integrin subunit. This work demonstrates that vWF is not only released into the systemic circulation, but is also leaked to the perivascular space/matrix. This local release and deposition of vWF is probably a sensitive and early marker of microvascular involvement in SSc pathogenesis. Local vWF release may play a role in platelet adhesion, aggregation, thrombogenesis and dermal connective tissue remodelling. In spite of some attempts towards compensatory angiogenesis in SSc, as evidenced by beta3 integrin subunit expression, it was evident that the angiogenic response was not able to prevent the development of hypovascularity during the advanced stages of the disease.