Effect of multi-nutrient insufficiency on markers of one carbon metabolism in young women: response to a methionine load.

BACKGROUND/OBJECTIVES: Multi-nutrient insufficiencies as a consequence of nutritional and economic factors are common in India and other developing countries. We have examined the impact of multi-nutrient insufficiency on markers of one carbon (1C) metabolism in the blood, and response to a methionine load in clinically healthy young women. SUBJECTS/METHODS: Young women from Pune, India (n=10) and Cleveland, USA (n=13) were studied. Blood samples were obtained in the basal state and following an oral methionine load (50 mg/kg of body weight in orange juice). Plasma concentrations of vitamin B12, folate and B6 were measured in the basal state. The effect of methionine load on the levels of methionine, total homocysteine, cysteine, glutathione and amino acids was examined. RESULTS: Indian women were significantly shorter and lighter compared with the American women and had lower plasma concentration of vitamins B12, folate and B6, essential amino acids and glutathione, but higher concentration of total homocysteine. The homocysteine response to methionine load was higher in Indian women. The plasma concentrations of glycine and serine increased in the Indian women after methionine (in juice) load. A significant negative correlation between plasma B6 and homocysteine (r= -0.70), and plasma folate and glycine and serine levels were observed in the Indian group (P<0.05) but not in the American group. CONCLUSIONS: Multi-nutrient insufficiency in the Indian women caused unique changes in markers of whole body protein and 1C metabolism. These data would be useful in developing nutrient intervention strategies.

Tissue engineered intervertebral disc repair in the pig using injectable polymers.

The intervertebral disc (IVD) has a central nucleus pulposus (NP) able to resist compressive loads and an outer annulus fibrosus which withstands tension and gives mechanical strength. The tissue engineering of a disc substitute represents a challenge from mechanical and biological (nutrition and transport) points of view. Two hyaluronan-derived polymeric substitute materials, HYAFF 120, an ester and HYADD 3, an amide were injected into the NP of the lumbar spine of female pigs (11.1 +/- 1.0 Kg) in which a nucleotomy had also been performed. Homologous bone marrow stem cells, obtained from the bone marrow three weeks before spinal surgery, were included in the HYADD 3 material (1x 10(6) cells/ml). Two lumbar discs were operated in each animal. Control discs received a nucleotomy only. The animals were killed after 6 weeks and the lumbar spines recovered for histopathological study. Nucleotomy resulted in loss of normal IVD structure with narrowing, fibrous tissue replacement and disruption of the bony end-plates (4/4). By contrast, both HYAFF 120 (4/4) and HYADD 3 (4/4) treatment prevented this change. The injected discs had a central NP-like region which had a close similarity to the normal biconvex structure and contained viable chondrocytes forming matrix like that of normal disc.

In vitro osteoblastic response to 30 vol% hydroxyapatite-polyethylene composite.

Hydroxyapatite-reinforced high-density polyethylene (HA-HDPE) composite, as a bone replacement material, has successfully been used clinically as middle ear prostheses and orbital floor implants. The aim of this study was to examine its in vitro biocompatibility in order to develop a further application, that is, as skull reconstruction implants. Human osteoblast cells isolated from femoral heads and crania were used to determine the biological response of the composites. HA-HDPE composites (30 vol %) with two grades of HA filler that had different surface morphologies were selected for this in vitro assessment. The results showed that HA-HDPE composite was bioactive and supported osteoblast attachment, proliferation, and differentiation. The composite with rough-surfaced HA filler demonstrated slightly better cellular response than the composite with smooth-surfaced HA filler. Although osteoblastic cells derived from skull showed an overall slower response compared with those from femoral heads, these in vitro results show that HA-HDPE composite potentially could be used as a skull implant.

The fundamentals of tissue engineering: new scaffolds.

The ability to regenerate new bone for skeletal use is a major clinical need. In this study, two novel porous calcium phosphate materials pure HA and biphasic HA/beta-Tricalcium phosphate (HA/beta -TCP) were evaluated as potential scaffolds for cell-seeded bone substitutes using human osteoblast-like cells (HOS) and primary human mesenchymal stem cells (hMSCs). A high rate of proliferation was observed on both scaffolds. A greater increase in alkaline phosphatase (ALP- an indicator of osteoblast differentiation) was observed on HA/beta -TCP compared to HA. This observation indicates that HA/TCP may play a role in inducing osteoblastic differentiation. Although further evaluation is required both materials show potential as innovative synthetic substitutes for tissue engineered scaffolds.

The fundamentals of tissue engineering: new scaffolds

The ability to regenerate new bone for skeletal use is a major clinical need. In this study, two novel porous calcium phosphate materials pure HA and biphasic HA/beta-Tricalcium phosphate (HA/beta -TCP) were evaluated as potential scaffolds for cell-seeded bone substitutes using human osteoblast-like cells (HOS) and primary human mesenchymal stem cells (hMSCs). A high rate of proliferation was observed on both scaffolds. A greater increase in alkaline phosphatase (ALP- an indicator of osteoblast differentiation) was observed on HA/beta -TCP compared to HA. This observation indicates that HA/TCP may play a role in inducing osteoblastic differentiation. Although further evaluation is required both materials show potential as innovative synthetic substitutes for tissue engineered scaffolds.

Biodegradable microspheres: a new delivery system for growth hormone.

A drug delivery system for biologically active agents targeted to specific cells could be used to improve tissue repair in orthopaedics. The system should be controllable and capable of drug release over an extended period of time. Biodegradable, membrane-moderated, monolithic microspheres for the controlled release of growth hormone (GH) were developed and the release of GH was monitored in vitro. Cross-linked gelatin microspheres were used as the vehicle, with the drug dispersed within the gelatin. The amount of GH released from the microspheres was increased following ultrasonication. The release of growth hormone was monitored in phosphate buffered saline and horse serum. Interestingly, a higher level of GH was detected in the phosphate buffered saline than in serum. In addition, both pH and enzyme-induced degradation had an effect on the swelling kinetics of the gelatin microspheres. The release of GH from the microspheres was diffusion controlled, during the time period studied.