Supplementation with green tea and oregano extracts on productive characteristics, blood metabolites, and antioxidant status of Jersey cows during the transition period.

Plant extracts have been recognized as beneficial to human health and have been evaluated as feed additive for domestic and companion animals. This study evaluated oregano and green tea extracts fed to Jersey cows from approximately 21 d before calving to 21 d after calving on milk production, milk composition, and blood metabolites as well as investigated immunological and antioxidant attributes. Twenty-four Jersey cows with 441 ±â€¯27 kg of BW, 3.5 ±â€¯0.3 of body condition score (BCS), and 2.7 ±â€¯1.8 lactations were selected at approximately 28 d before the expected parturition date and were randomly assigned to three treatments with eight cows each: without plant extracts in diet (control - CON), addition of 10 g per day of oregano extract (OR), and addition of 5 g per day of green tea extract (GT). Feed intake, BW, BCS, blood metabolites, hemogram as well as oxidative stress biomarkers were evaluated from approximately 3 weeks prepartum to 3 weeks postpartum (transition period) while milk production and composition were evaluated during the first 3 weeks of lactation. Plant extracts did not change BW, BCS, and DM intake (DMI) throughout the transition period, but OR increased in approximately 20% total digestive nutrients and metabolizable energy intake on days 15 and 16 postpartum compared with CON. In the prepartum, OR increased in 48% platelets count compared to the CON, while GT augmented in 142% eosinophils compared with CON. Oregano extract reduced the levels of reactive species in the erythrocytes in 40% during prepartum and postpartum compared with CON, while GT reduced its levels in 24 and 29% during prepartum and postpartum, respectively, when compared with CON. In the postpartum period, OR increased in 60% the carbonylated protein content compared with CON, while GT reduced in 45% the levels of reactive species in plasma compared with CON. During the postpartum, both extracts increased in 33% the concentration of reduced glutathione when compared with CON. Moreover, GT tended to decrease feed efficiency in 11% when compared with CON; OE reduced milk pH and somatic cell count when compared with CON. In conclusion, OE and GT did not expressively affect immunological attributes in blood but reduce some oxidative stress biomarkers without compromising productive traits of Jersey cows during the transition period.

Developmental programming: intrauterine caloric restriction promotes upregulation of mitochondrial sirtuin with mild effects on oxidative parameters in the ovaries and testes of offspring.

According to the developmental origins of health and disease (DOHaD) hypothesis, changes in the maternal environment are known to reprogram the metabolic response of offspring. Known for its redox modulation, caloric restriction extends the lifespan of some species, which contributes to diminished cellular damage. Little is known about the effects of gestational caloric restriction, in terms of antioxidant parameters and molecular mechanisms of action, on the reproductive organs of offspring. This study assessed the effects of moderate (20%) caloric restriction on redox status parameters, molecular expression of sirtuin (SIRT) 1 and SIRT3 and histopathological markers in the ovaries and testes of adult rats that were subjected to gestational caloric restriction. Although enzyme activity was increased, ovaries from female pups contained high levels of oxidants, whereas testes from male pups had decreased antioxidant enzyme defences, as evidenced by diminished glyoxalase I activity and reduced glutathione content. Expression of SIRT3, a deacetylase enzyme related to cellular bioenergetics, was increased in both ovaries and testes. Previous studies have suggested that, in ovaries, diminished antioxidant metabolism can lead to premature ovarian failure. Unfortunately, there is little information regarding the redox profile in the testis. This study is the first to assess the redox network in both ovaries and testes, suggesting that, although intrauterine caloric restriction improves molecular mechanisms, it has a negative effect on the antioxidant network and redox status of reproductive organs of young adult rats.

Transforming growth factor-beta1 incorporated during setting in calcium phosphate cement stimulates bone cell differentiation in vitro.

Growth stimulation of periimplant tissues by growth factors like transforming growth factor-beta1 (TGF-beta1) may increase the indication for and success of implant use. Calcium phosphate as a material for implants or for coating of implants is known for its good biologic interaction with bone. Therefore, calcium phosphate implants combined with TGF-beta1 might improve osseointegration. In this study we hypothesise that the addition of recombinant human TGF-beta1 (rhTGF-beta1) to calcium phosphate cement (CPC) affects the differentiation of bone cells growing on the cement layer. rhTGF-beta1 incorporated during setting in a CPC layer at 20 ng rhTGF-beta1/60 mg cement was found to be gradually released into tissue culturing medium leading to a 20% release after 24 h. Two cell populations were obtained from collagenase-treated fragments of adult rat long bones: preosteoblastic cells, which were released by the collagenase treatment, and osteoblastic cells, which grew from the collagenase-stripped bone fragments. Both cell populations were tested for their osteoblastic characteristic phenotype by measuring their alkaline phosphatase (ALP) activity after vitamin D treatment and cyclic AMP after parathyroid hormone stimulation. After preculture the cells were plated on a layer of CPC containing 0 (control), 10, or 20 ng rhTGF-beta1/60 mg CPC. Bone cell differentiation was analyzed after 10 days by measuring the ALP activity, as well as the protein content of the cell layer. Incorporation of rhTGF-beta1 in the CPC did not change the ALP activity in osteoblastic cells, but a significant (analyzed by multivariate analysis of variance) increase was observed in preosteoblastic cells. Incorporation of 10 ng of rhTGF-beta1 in 60 mg of CPC increased the ALP activity in preosteoblastic cells by threefold and 20 ng rhTGF-beta1/60 mg CPC increased it by fivefold. The total protein content was not affected by rhTGF-beta1 in either of the cell populations. We conclude that rhTGF-beta1 incorporated during setting in CPC stimulates the differentiation of preosteoblastic cells in vitro. These results provide a basis for further studies on the use of this combination as an implant material in vivo.

Fatigue behavior of calcium phosphate coatings with different stability under dry and wet conditions.

To obtain stable plasma sprayed calcium phosphate coatings, coatings with a high crystallinity and low solubility were developed. However, stability of ceramic coatings is also influenced by their fatigue resistance. Recently, fatigue failure was proposed to explain coating detachment from implants under loaded conditions. Therefore, plasma-sprayed calcium phosphate coatings with different crystallinity were investigated in vitro for fatigue failure. An amorphous and a crystalline hydroxylapatite coating (AHA and CHA) and a highly crystalline fluorapatite coating (FA) were subjected to cyclic load tests, both in dry conditions and in simulated body fluid (SBF). The results in SBF revealed that the crystalline CHA and FA coating detached completely at the highest stressed middle section of the bar. The FA coating delaminated earlier than the CHA coating. The amorphous AHA coating showed only partial coating loss at the completion of the test. Tests in dry conditions did not reveal any change in the coatings tested. These results suggest a relation between crystallinity of apatite coatings and their failure due to fatigue: high crystallinity coatings demonstrate earlier and more complete fatigue failure than the amorphous apatite coatings. It can be concluded that coating stability is not determined solely by static dissolution, but by fatigue failure as well.

Hepatic arterial pulsatility index in cirrhosis: correlation with portal pressure.

BACKGROUND/AIMS: Determination of the pulsatility index by means of duplex sonography provides the opportunity to evaluate the vascular resistance of the hepatic artery noninvasively. The aim of this study was to investigate the relationship between the hepatic arterial pulsatility index and the hepatic venous pressure gradient in cirrhosis. METHODS: In 50 patients with cirrhosis, hepatic venous pressure gradient was determined in the fasting state. Immediately thereafter, hepatic arterial pulsatility index and portal blood flow velocity were measured by duplex sonography with no knowledge of hepatic venous pressure values. In addition, the duplex parameters were determined in 20 controls. RESULTS: Hepatic arterial pulsatility index was significantly higher in patients with cirrhosis than in controls (0.92+/-0.1 vs. 1.14+/-0.18; p<0.001) and directly correlated with the hepatic venous pressure gradient (r = 0.7; p<0.001). Furthermore, weak correlations were found between hepatic arterial pulsatility index and Child-Pugh score (r = 0.49; p<0.01) and between portal blood flow velocity and hepatic venous pressure gradient (r = -0.48; p<0.01). CONCLUSION: In cirrhosis the hepatic arterial vascular resistance seems to increase parallel to the rise of the portal pressure. Therefore, duplex sonographic determination of the hepatic arterial pulsatility index may contribute to the noninvasive evaluation of portal hypertension.

Bone tissue response to biodegradable polymers used for intra medullary fracture fixation: a long-term in vivo study in sheep femora.

Since the degradability of the synthetic polymers used for fracture fixation is still unclear, a research project with biodegradable interlocking nails with a longterm implantation period has been started. In 21 female sheep a complete mid-shaft osteotomy of the left femur was performed to mimic a fracture of the femoral shaft. For the fixation, an intramedullary stainless-steel interlocking nail, a PLA rod or a PLA/PGA rod was used. After 30 months of implantation the histological results of these three materials were examined. In contrast to most reports the degradation rate of both polymers was much lower than the suggested ultimate period of two years. Even the tissue response was more pronounced than expected and this reaction can imply certain risks for repulsion. One can conclude that the volume quantity of polymeric implant in the bony tissue must be reduced if possible to avoid severe foreign body responses. The immunologic responses and the clinical consequences need more studies. The degradation behavior of the polymer is still not under control.

Effect of losartan, an angiotensin II receptor antagonist, on portal pressure in cirrhosis.

Administration of angiotensin II causes an increase in portal pressure, and plasma concentration of angiotensin II is elevated in patients with cirrhosis, suggesting that angiotensin II may be involved in the pathogenesis of portal hypertension in cirrhosis. We evaluated the effect of the orally active angiotensin II receptor antagonist, losartan, on portal pressure in patients with cirrhosis and portal hypertension. Thirty patients with severe (hepatic venous pressure gradient [HVPG] >/= 20 mm Hg) and 15 patients with moderate (HVPG < 20 mm Hg) portal hypertension at baseline measurement were treated with an oral dose of 25 mg losartan once daily for 1 week and compared with 15 (HVPG >/= 20 mm Hg) and 10 (HVPG < 20 mm Hg), respectively, cirrhotic controls. On the seventh day, HVPG was determined again, and blood pressure, heart rate, body weight, and parameters of liver and kidney function were recorded. Losartan induced a significant (P <.001) decrease of HVPG in the patients with severe (-46.8% +/- 15.5%) and moderate (-44.1% +/- 14.7%) portal hypertension, while no significant change was seen in the controls. Losartan caused a slight but significant (P <.01) fall in mean arterial blood pressure (-3.1 +/- 5.0 and -3.5 +/- 4.3 mm Hg, respectively). One patient treated with losartan had a short symptomatic hypotensive reaction after the first dose of losartan that did not recur despite continued treatment. No deterioration of liver or kidney function was observed. The present study indicates that angiotensin II blockade with orally administered losartan is safe and highly effective in the treatment of portal hypertension.

Cortical bone ingrowth in growth hormone-loaded grooved implants with calcium phosphate coatings in goat femurs.

Dental implants are successfully used for tissue-integrated protheses, but the long-term survival in the maxilla is shorter than in the mandible [Cune MS, Thesis, University of Utrecht, 1993; Jaffin RA, Berman CL. J Periodontol 1991;62:2-4]. However, by adding growth hormone at implantation, increased bone apposition may be expected, since it is known that growth hormone has a stimulating effect on the differentiation and proliferation of osteoblastic cells [Ernst M, Froesch ER. Biochem Biophys Res Commun 1988;151:142-47; Scheven BAA et al. Growth Regul 1991;1:160-67; Stracke H et al. Acta Endocrinol 1984;107:16-24]. We studied bone ingrowth and bone contact of grooved implants impregnated with growth hormone in the cortex of femurs of female goats. We compared the effect of growth hormone on grooved implants with or without calcium phosphate coatings at the bottom of the grooves. The coatings used were hydroxyapatite, fluorapatite and heat-treated hydroxyapatite. The implants had both small and large grooves. The implants were positioned in the cortex of one femur and were treated with recombinant human growth hormone, while the implants on the opposite femur served as controls. After 6 weeks, the implants and surrounding tissues were dissected and evaluated histomorphologically and morphometrically by light microscopy. The bone ingrowth and the bone contact in the grooves were quantified by digital image analysis. Calcium phosphate coating at the bottom of the grooves resulted in a significant increase of bone ingrowth and bone contact. Small grooves had significantly more bone ingrowth and bone contact than the larger grooves. However, all implants impregnated with growth hormone showed inhibition of bone contact and bone ingrowth. We conclude that recombinant human growth hormone inhibits bone formation in the grooves coated with calcium phosphate. Without the addition of growth hormone, the calcium phosphate coatings improved bone ingrowth and bone contact in the grooves. Further studies are required to determine whether growth hormone could also possibly act as a bone growth promoting factor in these implants.

Healing of large (2 mm) gaps around calcium phosphate-coated bone implants: a study in goats with a follow-up of 6 months.

Plasma-sprayed hydroxylapatite (HA) coatings are known for their ability to demonstrate osseointegration with bone. Recently it was found that the amount of bone apposition was strongly reduced 6 weeks after implantation in a goat model if gaps of two millimeters between bone and apatite coating existed. Stability of the apatite coatings examined did not influence the gap-healing ability. This study investigated whether a longer follow-up period of 24 weeks would be sufficient for the restoration of bone apposition on apatite coatings in an identical surgical model with 2 mm gaps, and whether bone apposition on the apatite coatings is influenced by the coating stability. Three coatings were investigated: 25-30% crystalline HA (aHA), 60-63% crystalline HA (cHA), and 85-90% crystalline fluorapatite (FA). Uncoated Ti-6A1-4V implants were used as controls. Implants were inserted in the femoral condyles of both femora of eight goats. Each goat received four implants. Histology revealed that bone formation on each of the apatite coatings remained low and did not increase with an extended follow-up period of 24 weeks. The coatings showed significantly (P < 0.01) more bone contact than the uncoated control implants. The three different coatings did not show significant differences in bone apposition. The aHA coating in most cases had disappeared completely after 24 weeks. Despite the disappearance of the aHA coating, bone contact was seen on the substrate surface without fibrous tissue interposition. The cHA coating showed minor signs of degradation while the FA coatings showed no visible degradation. It is concluded that non-press-fit implantation of apatite-coated implants leads to more bone apposition as compared to uncoated Ti-6A1-4V implants. However, it is suggested by these results that the upper limit of gaps around apatite implants is 2 millimeters in a non-weight-bearing model in goats. Bone apposition will not increase by extending the follow-up period more than six weeks, nor will it be altering the stability of the apatite coatings used.

Biodegradable interlocking nails for fracture fixation.

Serious problems such as stress shielding, allergic reactions, and corrosion are associated with the use of metallic fracture fixation devices in fractured long bones. Metal implants often are removed during a second retrieval operation after fracture healing has completed. A biocompatible implant that degrades slowly during implantation would obviate the need for a second operation and save the patient from considerable physical, psychologic, and financial discomfort. The biodegradable implant must provide the fractured limb sufficient support for a certain time, allowing early loading. A gradual transfer of load from the biodegradable implant to the bone would result in a better product of bone healing and avoid stress shielding. In an animal model using adult sheep, two types of biodegradable polymer interlocking nails were tested in comparison with a stainless steel interlocking nail. Fracture healing, mechanical properties of the bones, degradation behavior in vivo and in vitro, and tissue response were monitored during a 2 1/2-year followup study. To detect shifts in acid base relations caused by the release of acid compounds, pH measurements were performed. Fracture healing was unimpaired, and the mechanical test results of all three groups were excellent. Histologic analysis showed a mild inflammatory response, but no pH shifts were observed. The results of this study justify additional research on these promising materials.

Healing of gaps around calcium phosphate-coated implants in trabecular bone of the goat.

Hydroxylapatite coatings are under clinical investigation in orthopaedics and dentistry. Bone formation on apatite coatings in the presence of gaps is important for clinical applications. The importance of the stability of the coating is not known at present. By varying the plasma-spray parameters, and by the addition of fluoride, the crystallinity and stability of calcium phosphates can be changed. It is suggested that bone formation is enhanced by dissolution of the apatite coating. We studied apatite coatings of varying stability with regard to their gap-healing characteristics, and we examined what the maximum gap would be that can be bridged if a coating is applied. Ti-6A1-4V implants coated with 62% crystalline hydroxylapatite, 30% crystalline hydroxylapatite or fluorapatite, or noncoated Ti-6A1-4V were implanted in 16 goats. The implants were surrounded by gaps of 1 or 2 mm, and the follow-up period was 6 weeks. Histological examination and histometry revealed that gaps of 1 mm can be bridged by bone if an apatite coating is applied. However, only a minimal amount of bone contact was seen on the apatite coatings with 2 mm gaps. Uncoated implants demonstrated no bone contact at all. Among the three different coatings there were no differences in gap healing. It can be concluded that in the goat, gaps of 2 or more mm between coated implants and host bone tissue inhibit bone deposition on the coating (p < 0.05), but the stability of the coating does not influence gap-healing characteristics.

In vivo study of calcium phosphate cements: implantation of an alpha-tricalcium phosphate/dicalcium phosphate dibasic/tetracalcium phosphate monoxide cement paste.

alpha-Tricalcium phosphate (alpha-TCP)/dicalcium phosphate dibasic (DCPD)/tetracalcium phosphate monoxide (TeCP) cement was implanted in paste form into soft tissue (rate subcutaneous sockets) and bone tissue (defects in rabbit mandibles) to evaluate the setting behaviour of the cement and tissue responses to the cement. A histological study of the soft tissue implants revealed thin fibrous capsule formation, the appearance of multinucleated giant cells on and close to the cement surface, and small clusters of the cement near the main part of the set cement which were formed by the migration of the paste while setting. X-ray diffraction (XRD) analysis of the implanted cement showed peaks for hydroxyapatite (HA) which increased as the implant period increased. Histology and microradiography of the bone tissue implants showed well-set cement without migration, active bone formation around the cement and direct bone union to it. However, the cement disappeared from the implant site in 4 of 16 specimens where intense bleeding seemed to wash away the implants while setting. From the results of the present study, we concluded that the cement is well tolerated, especially by bone tissue. This may be related to the fact that the cement sets producing HA. The cement is a promising material as a bone substitute; however, there is a problem of migration while setting in soft tissue and of exclusion from the bone defects by intense bleeding.

Behavior of calcium phosphate coatings with different chemistries in bone.

Calcium phosphate ceramic coatings with a hydroxyapatite chemistry applied on the surface of dental implants eliminate the need for initial mechanical retention and decrease the time necessary for bonding the implants to the bone. Hydroxyapatite-coated implants retrieved from patients were found to be compatible and to have bonded strongly to the bone, but the coatings showed thinning because of partial or total loss of coating material. This study compared the behavior in bone of newly developed fluorapatite and heat-treated hydroxyapatite coatings, with the clinically used hydroxyapatite coatings used as controls in experimental studies in dogs. The biologic responses to fluorapatite and heat-treated hydroxyapatite coatings were the same as those to hydroxyapatite coatings, and bone condensation around all coatings was histologically evident. However, the coating thickness of the fluorapatite and heat-treated hydroxyapatite coatings remained stable with only minor changes during the observation period of 24 months.

The burst phenomenon, an animal model simulating the long-term tissue response on PLLA interlocking nails.

In this in vivo study, low-molecular-weight as-polymerized PLLA powder was placed in the medullary cavity of a porcine femur in order to study the tissue reaction on predegradated PLLA. An attempt was made to simulate the long-term degradation of a large PLLA implant. This phase can be characterized by the release of PLLA particles and acid compounds from a heterogeneously degrading PLLA implant into the surrounding tissues. The clinical consequences and tissue response were studied. The clinical recovery of the experimental animals was favorable. No signs of clinical inflammation could be detected during the eight-week follow-up period. Histological analysis of the bone/PLLA interface showed signs of a mild inflammatory tissue response.

Comparison of the release of growth hormone from hydroxyapatite, heat-treated hydroxyapatite, and fluoroapatite coatings on titanium.

Hydroxyapatite (HA), heat-treated hydroxyapatite (HAH), and fluorapatite (FA) coatings on titanium were loaded with human growth hormone (GH), and the subsequent release was monitored in vitro. The amount of GH released was significantly increased from the HA coating that had received a post-plasma-spraying heat treatment prior to incorporation of the growth hormone. Scanning electron microscopy was used to assess the surfaces of the ceramic coatings prior and postincubation with GH. Surface changes were observed on the HA and HAH coatings but not on the FA coatings after incubation with GH. Osteoblast-like cells were grown on the coatings and maintained in culture. Scanning electron microscopy was used to study the morphology of the cells and the interaction of the cells with the ceramic coatings. We used thymidine uptake and DNA content to determine the relative rates of cell division on the different coatings; the optimum rate of cell proliferation was observed on the HAH coating loaded with 0.1 IU/mL GH.

Effect of sintering temperature on silica gels and their bone bonding ability.

Silica gels gave rise to apatite precipitation onto their surface depending on the sintering temperature. Silica gels prepared at different sintering temperatures were studied in vivo in cortical bone for their bone bonding ability in relation to their apatite precipitation. From the results we conclude that the sintering temperature influenced the stability of the material. Sintering at the lower temperatures of 400 and 600 degrees C allow the silica gels to be degraded more easily, while gels treated at 900 and 1000 degrees C were more stable. The more stable gels showed some bone bonding, while the degraded gels evoked a high cellular reaction of giant cells and lymphocytes.

A model for the evaluation of mandibular bone response to implant materials.

An in vivo animal (goat) mandibular bone model has been evaluated for the cortical and trabecular bone response to biomaterials with different elastic modulus and/or different surfaces. The effect of the elastic modulus on the bone formation was also studied. The difficulties encountered in orientation of the implants probably could have been surmounted if pre-operative radiographs had been available. It was also established that there is a large variance in the amount of trabecular and cortical bone in the mandible of the goat 'bone model'.

In vivo calcium phosphate formation induced by sol-gel-prepared silica.

Implantation with plugs made of a porous sol-gel-prepared silica into the femurs of goats demonstrated that a calcium phosphate was formed both on the silica plugs and within the pores inside the silica plugs 12 weeks postoperatively. This observation indicates that a highly hydrated silica surface is effectively catalytic for calcium phosphate nucleation. Calcification can be triggered in physiologic solution under stimulation of the silica gel. A high level of silicon in the uncalcified osteoid region of young bone is thus thought to provide a number of SiOH groups for initiating calcium phosphate formation. Our results provide some information about the mechanism of calcium phosphate mineralization in higher animals. We believe that heterogeneous nucleation of apatite can be induced from metastable calcium phosphate solutions including physiologic fluids on those specific surfaces of materials, where there are abundant acidic OH groups.

Tissue reaction on PLLA versus stainless steel interlocking nails for fracture fixation: an animal study.

In order to develop a biodegradable interlocking nail for fracture fixation, polylactic acid (PLLA) rods were implanted in the femoral bone of young pigs. A midshaft osteotomy was performed in order to mimic a fracture. The tissue response to PLLA rods versus stainless steel rods was studied after 1 and 3 months of implantation. Fracture-healing characteristics and chemical and mechanical properties of the PLLA rods were studied after explantation. Mechanical properties deteriorated during implantation and chemical properties changed significantly in 1 and 3 months of implantation. PLLA and stainless steel induced a similar tissue reaction during the studied implantation time of 3 months.