In vivo mechanical loading modulates insulin-like growth factor binding protein-2 gene expression in rat osteocytes.

Mechanical stimulation is essential for maintaining skeletal integrity. Mechanosensitive osteocytes are important during the osteogenic response. The growth hormone-insulin-like growth factor (GH-IGF) axis plays a key role during regulation of bone formation and remodeling. Insulin-like growth factor binding proteins (IGFBPs) are able to modulate IGF activity. The aim of this study was to characterize the role of IGFBP-2 in the translation of mechanical stimuli into bone formation locally in rat tibiae. Female Wistar rats were assigned to three groups (n = 5): load, sham, and control. The four-point bending model was used to induce a single period of mechanical loading on the tibial shaft. The effect on IGFBP-2 mRNA expression 6 hours after stimulation was determined with nonradioactive in situ hybridization on decalcified tibial sections. Endogenous IGFBP-2 mRNA was expressed in trabecular and cortical osteoblasts, some trabecular and subendocortical osteocytes, intracortical endothelial cells of blood vessels, and periosteum. Megakaryocytes, macrophages, and myeloid cells also expressed IGFBP-2 mRNA. Loading and sham loading did not affect IGFBP-2 mRNA expression in osteoblasts, bone marrow cells, and chondrocytes. An increase of IGFBP-2 mRNA-positive osteocytes was shown in loaded (1.68-fold) and sham-loaded (1.35-fold) endocortical tibial shaft. In conclusion, 6 hours after a single loading session, the number of IGFBP-2 mRNA-expressing osteocytes at the endosteal side of the shaft and inner lamellae was increased in squeezed and bended tibiae. Mechanical stimulation modulates IGFBP-2 mRNA expression in endocortical osteocytes. We suggest that IGFBP-2 plays a role in the lamellar bone formation process.

Additional weight bearing during exercise and estrogen in the rat: the effect on bone mass, turnover, and structure.

Mechanical loading and estrogen play important roles in bone homeostasis. The aim of this study was to evaluate the effects of mechanical loading on trabecular bone in the proximal femur of ovariectomized rats. We hypothesized that mechanical loading suppresses bone resorption and increases bone formation, which differs from the suppressive effects of estrogen on both resorption and formation. Furthermore, we expected to find changes in trabecular architecture elicited by the effects of mechanical loading and estrogen deficiency. Sixty female Wistar rats, 12 weeks old, were assigned to either the sedentary groups sham surgery (SED), ovariectomy (SED+OVX), and ovariectomy with estrogen replacement (SED+OVX+E2) or to the exercise groups EX, EX+OVX, EX+OVX+E2. Following ovariectomy, 5 microg 17beta-estradiol was given once weekly to the estrogen replacement groups. Exercise consisted of running with a backpack (load +/-20% of body weight) for 15 minutes/day, 5 days/week, for 19 weeks. Dual-energy X-ray absorptiometry (DXA) scans were performed before (T0), during (T6), and after (T19) the exercise period to obtain bone mineral content (BMC) and bone mineral density (BMD) data. After the exercise program, all rats were killed and right and left femora were dissected and prepared for micro-CT scanning and histomorphometric analysis of the proximal femoral metaphysis. After 19 weeks, increases in BMC (P = 0.010) and BMD (P = 0.031) were significant. At T19, mechanical loading had a significant effect on BMC (P = 0.025) and BMD (P = 0.010), and an interaction between mechanical loading and estrogen (P = 0.023) was observed. Bone volume and trabecular number decreased significantly after ovariectomy, while trabecular separation, mineralizing surface, bone formation rate, osteoclast surface, degree of anisotropy, and structure model index increased significantly after ovariectomy (P < 0.05). Trabecular bone turnover and structural parameters in the proximal femur were not affected by exercise. Estrogen deficiency resulted in a less dense and more oriented trabecular bone structure with increased marrow cavity and a decreased number of trabeculae. In conclusion, mechanical loading has beneficial effects on BMC and BMD of the ovariectomized rat. This indicates that the load in the backpack was high enough to elicit an osteogenic response sufficient to compensate for the ovariectomy-induced bone loss. The results confirm that estrogen suppresses both bone resorption and bone formation in the proximal metaphysis in the femoral head of our rat-with-backpack model. The effects of mechanical loading on the trabecular bone of the femoral head were not significant. This study suggests that the effect of mechanical loading in the rat-with-backpack model mainly occurs at cortical bone sites.

Histological observations on biopsies harvested following sinus floor elevation using a bioactive glass material of narrow size range.

We evaluated the bone augmenting capacity of bioactive glass particles, size range 300-355 microns (BG-particles), in human sinus floor elevations using histomorphometrical methods. A total of 10 patients underwent bilateral grafting, using a 1:1 mixture of autogenous bone particles (from iliac crest) and BG-particles at one side (experimental side), and bone particles only at the other side (control side, split mouth design). A total of 72 bone biopsies were taken at the time of fixture installation; that is, 3 patients at 4 months, 3 at 5 months and 3 at 6 months after grafting and 1 patient at 16 months (when she presented again). In each case 6 biopsies were taken, 3 left and 3 right. Histomorphometry showed that in grafts at control sides, trabecular bone was present after 4 months, comprising almost 41% of the tissue volume. This bone contained viable osteocytes and was of mature lamellar type and showed a mature histological appearance. Bone volume continued to increase slightly, to 42% at 5 months, 44% at 6 months and 45% at 16 months. The graft volume at experimental sides consisted at 4 months for 28% of woven and some lamellar bone, and increased to 35% at 5 months and 38% at 6 months, when mainly lamellar bone was found. At 16 months a lamellar bone volume of 45% was found. The BG-particles transformed and became excavated with time, starting at 4 months, and their centers gradually filled with bone tissue. All BG-particles had disappeared by resorption at 16 months after grafting and had been replaced by bone tissue. Parameters of bone turnover (% osteoid surface, % resorption surface, mineral apposition rate as measured by tetracycline labeling) indicated that bone remodeling was very active at both sides, during more than 6 months, despite the mature histological appearance of the bone tissue. From these histological observations, we conclude that a 1:1 mixture of autogenous bone/BG-particles seems a promising alternative to autogenous bone only, when low amounts of bone tissue are available for sinus augmentation.

The development of daughter sporocysts inside the mother sporocyst of Schistosoma mansoni with special reference to the ultrastructure of the body wall.

The development of the mother sporocyst and the differentiation of the daughter sporocyst of Schistosoma mansoni in Biomphalaria pfeifferi are described. The tegumental structure of the mother sporocyst, consisting of an outer layer connected to internally situated nucleated cell bodies, forms extensions which enwrap the germinal cells. The parenchyma cells, in which the germinal cells were embedded before, degenerate. When daughter sporocyst embryos develop from germinal cells they are enveloped by a primitive epithelium which is formed by fusion of the extensions of the tegumental structure of the mother sporocyst. Somatic cells located peripherally in the developing daughter sporocyst expand and coalesce beneath the primitive epithelium to form the future outer layer of the tegumental structure of the daughter sporocyst. The primitive epithelium degenerates, the newly-formed layer looses its nuclei, and becomes connected to internally situated nucleated cell bodies. Further developments in the tegumental structure of the daughter sporocyst include the formation of microvillus-like projections, a surface coat, spines, and a basement membrane.

Ultrastructural changes in the body wall of Schistosoma mansoni during the transformation of the miracidium into the mother sporocyst in the snail host Biomphalaria pfeifferi.

The ultrastructure of the body wall of the free miracidium of Schistosoma mansoni and the changes occurring within 48 h after penetration into the intermediate host Biomphalaria pfeifferi are described. Within 2 h after penetration the ciliated plates are shed into the haemolymph of the snail and phagocytized by amoebocytes. At the same time the narrow ridges between the plates of the free miracidium expand to form the continuous outer layer of the sporocyst. Within 48 h the entire tegumental structure, consisting of a thin outer layer, connected with sunken nucleated areas, develops to its full extent. The observations are compared with those on Fasciola hepatica.

The development of the primitive epithelium and true tegument in the cercaria of Schistosoma mansoni.

The formation of the final cercarial tegument of Schistosoma mansoni is preceded by that of a so-called primitive epithelium. The primitive epithelium is derived from the tegument of the daughter sporocyst. The final cercarial tegument is formed from peripherally located somatic cells of the cercarial embryo, which expand and coalesce beneath the primitive epithelium. The primitive epithelium degenerates and disappears. The ultrastructure of both epithelia in the course of the development of the cercaria is described in detail. Possible functions are discussed.