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Biomedical subjects

Martin Kramer

Publications and source records attributed to Martin Kramer.

14 recordsLinked to original sources
PubMed2006

Platelet-rich plasma improves expansion of human mesenchymal stem cells and retains differentiation capacity and in vivo bone formation in calcium phosphate ceramics.

INTRODUCTION: Mesenchymal stem cells (MSC) applied to bone substitution materials can improve bone healing. Bone formation in biocomposites is highly dependent on the kind of biomaterial, its pre-treatment and the applied cells. Potentially immunogenic or infectious supplements such as fetal calf serum (FCS) should be avoided in cell expansion media. Therefore, we developed an expansion protocol free of xenogenic supplements. Cells expanded with two different media were tested on distinct biomaterials for their bone formation capacity after ectopic implantation in vivo, as well as for their growth rate and differentiation capacity in vitro. METHODS: MSC of six donors were expanded with cell expansion medium containing FCS (2%) or platelet-rich plasma (PRP, 3%). Their growth rate and osteogenic, adipogenic and chondrogenic differentiation capacity were compared in vitro. For the in vivo bone formation assay, expanded cells (2 x 105 or 2 x 106) were seeded on calcium-deficient hydroxyapatite (CDHA; n = 12) and on beta-tricalcium phosphate (beta-TCP; n = 12) blocks, which had been coated with either fibronectin or human serum. They were then implanted subcutaneously in severe combined immunodeficient mice (SCID), harvested after 8 weeks and analysed by histology. Bone formation was assessed by a semi-quantitative bone score, after toluidine blue and alizarin red staining. Human cells were detected by an in situ hybridisation for human-specific alu sequences. RESULTS: PRP-supplemented expansion medium yielded two-fold higher cell numbers compared to medium with FCS (P = 0.046) after 3 weeks (four passages) and retained a similar capacity to differentiate towards the osteogenic, chondrogenic and adipogenic lineage. In vivo bone formation was equal for cells expanded with PRP and FCS and depended on the specific surface area of the carrier. CDHA (specific surface area (SSA) 48 m2/g) showed a significantly better bone formation in deep layers (P = 0.005) than beta-TCP (SSA 0.5 m2/g). Fibronectin-coating of the ceramics was slightly superior to coating with human serum (P = 0.045). CONCLUSIONS: The replacement of FCS by PRP eliminated risks connected with the use of xenogeneic supplements. It improved expansion of MSC and retained their differentiation and in vivo bone formation capacity in a setting adaptable to autogenous use.

Animals↗
PubMed2006

Influence of platelet-rich plasma on osteogenic differentiation of mesenchymal stem cells and ectopic bone formation in calcium phosphate ceramics.

Platelet-rich plasma (PRP) contains a mixture of growth factors that play an important role in wound and fracture healing. While PRP enhanced bone formation by autogenous cancellous bone grafts, its influence in combination with different bone substitutes remained unknown. This study evaluated the effect of PRP on osteogenic differentiation and ectopic bone formation of human mesenchymal stem cells (MSC) in distinct resorbable calcium phosphate ceramics. Calcium-deficient hydroxyapatite (CDHA) blocks with a large specific surface area (48 m2/g) and beta-tricalcium phosphate (beta-TCP) with a low specific surface area (<0.5 m2/g) were loaded with 2 x 10(5) bone marrow-derived MSC. Half of the specimens were treated with 5-fold concentrated PRP. Biocomposites were implanted subcutaneously into SCID mice or kept under osteogenic culture conditions for 2 weeks before implantation. The addition of PRP increased the specific alkaline phosphatase (ALP) activity (p = 0.012) in undifferentiated MSC/CDHA composites but not in MSC/beta-TCP composites. Osteogenic preinduction was ineffective for CDHA and reduced ALP activity of beta-TCP composites significantly at explantation. Ectopic bone formation was stronger in MSC/CDHA (7/32) compared to MSC/beta-TCP (2/30) composites, but no influence of PRP was evident. In conclusion, the effect of PRP depended on the type of ceramic and the differentiation status of the MSC, and enhanced ALP activity of MSC on the high surface scaffold CDHA only, but PRP did not improve osteogenesis in our setting.

Aged↗
PubMed2006

Evaluation of nonunion fractures in dogs by use of B-mode ultrasonography, power Doppler ultrasonography, radiography, and histologic examination.

OBJECTIVE: To investigate the use of ultrasonography to assess nonunion of fractures in dogs and to compare results of ultrasonography, radiography, and histologic examination. SAMPLE POPULATION: 8 nonunion fractures in 6 dogs (1 each in 5 dogs and 3 in 1 dog); dogs ranged from 7 to 94 months of age and weighed 6 to 30 kg. PROCEDURES: Diagnostic assessment consisted of complete clinical and orthopedic examinations, radiography, B-mode (brightness mode) ultrasonography, and power Doppler ultrasonography. Biopsy samples were obtained during surgery for histologic examination. They were stained with H&E and immunolabeled by use of anti-CD31 antibodies. Correlations of power Doppler score, power Doppler count, vessel area, and radiographic prediction with the mean number of vessels counted per hpf were derived. RESULTS: Radiographically, 7 of 8 nonunion fractures were diagnosed as atrophic and were therefore estimated to be nonviable. Vascularity of nonunion fractures during power Doppler ultrasonography ranged from nonvascularized to highly vascularized. Absolute vessel count during histologic examination ranged from 0 to 63 vessels/hpf; 5 nonunion sites had a mean count of > 10 vessels/hpf. Vascularity during power Doppler ultrasonography was highly correlated with the number of vessels per hpf, whereas the correlation between the radiographic assessment and histologic evaluation was low. CONCLUSIONS AND CLINICAL RELEVANCE: Radiographic prediction of the viability of nonunion fractures underestimated the histologically assessed vascularity of the tissue. Power Doppler ultrasonography provided a more accurate estimation of the viability of the tissue and therefore the necessity for debridement and autografts during revision surgery.

Animals↗
PubMed2005

Reduced cord blood immune effector-cell responsiveness mediated by CD4+ cells induced in utero as a consequence of placental Plasmodium falciparum infection.

To determine mechanisms of neonatal parasite antigen (Ag)-specific immune suppression associated with placental Plasmodium falciparum infection, we isolated cord blood mononuclear cells (CBMCs) from Gabonese neonates born to mothers with differing histories of P. falciparum infection and performed ex vivo and in vitro studies to evaluate immune regulatory activity. We found increased ex vivo percentages of CD4(+)CD25(hi) and CD4(+)CD25(+)CTLA-4(+) cells and increased interleukin (IL)-10 responses to parasite Ag in vitro in CBMCs from neonates born to mothers with placental P. falciparum infection at delivery. Depleting CBMCs of CD4(+)CD25(+) cells before cell culture led to the abrogation of parasite Ag-specific IL-10 responses, to enhanced interferon- gamma responses, and to enhanced expression of CD25 on CD8(+) T cells and of major histocompatibility complex class I and II on monocytes. These data demonstrate that parasite Ag-specific CD4(+) regulatory cells are generated in utero as a consequence of placental P. falciparum infection.

Adult↗
PubMed2005

Clinical and parasitological characteristics of puerperal malaria.

BACKGROUND: Women with semi-immunity to malaria who live in regions where the disease is endemic are at increased risk for more frequent and severe episodes of malaria during pregnancy. Recent findings indicate that this increased risk might persist beyond delivery, but the underlying mechanisms for this change in risk are poorly understood. METHODS: One hundred fifty women were included in a cohort study in Lambaréné, Gabon, and were actively followed up weekly for 10 weeks after delivery, as were nonpregnant control women who had been matched to them by location and age. Parasites in samples of placenta and blood were genotyped by use of polymerase chain reaction amplification of the merozoite surface antigen 2 gene and the subtelomeric variable open reading frame gene of Plasmodium falciparum. RESULTS: Eleven puerperal women had cases of clinical malaria, compared with 1 control woman (rate ratio, 9.8; P=.006). Eighteen puerperal women had P. falciparum parasitemia, compared with 6 control women (rate ratio, 2.7; P=.03). Five of 16 puerperal women (31%) with parasitemia on follow-up had identical parasites in their placentas and blood, and 11 of these cases (69%) were the result of reinfection. Puerperal women remained at equal risk for the development of parasitemia throughout the first 10 weeks after delivery. Use of bed nets, use of chloroquine prophylaxis during pregnancy, presence of malaria episodes during pregnancy, gravidity, and age were not associated with the acquisition of parasitemia during follow-up. CONCLUSIONS: Compared with nonpregnant women, puerperal women have a considerably increased risk for the development of malaria and/or parasitemia. This increased risk is caused both by the recurrence of P. falciparum parasitemia and by the increased susceptibility to new infections, although the latter plays a more significant role.

Adult↗
PubMed2005

IFN-gamma and IL-10 mediate parasite-specific immune responses of cord blood cells induced by pregnancy-associated Plasmodium falciparum malaria.

Available evidence suggests that immune cells from neonates born to mothers with placental Plasmodium falciparum (Pf) infection are sensitized to parasite Ag in utero but have reduced ability to generate protective Th1 responses. In this study, we detected Pf Ag-specific IFN-gamma(+) T cells in cord blood from human neonates whose mothers had received treatment for malaria or who had active placental Pf infection at delivery, with responses being significantly reduced in the latter group. Active placental malaria at delivery was also associated with reduced expression of monocyte MHC class I and II in vivo and following short term in vitro coculture with Pf Ag compared with levels seen in neonates whose mothers had received treatment during pregnancy. Given that APC activation and Th1 responses are driven in part by IFN-gamma and down-regulated by IL-10, we examined the role of these cytokines in modulating the Pf Ag-specific immune responses in cord blood samples. Exogenous recombinant human IFN-gamma and neutralizing anti-human IL-10 enhanced T cell IFN-gamma production, whereas recombinant human IFN-gamma also restored MHC class I and II expression on monocytes from cord blood mononuclear cells cocultured with Pf Ag. Accordingly, active placental malaria at delivery was associated with increased frequencies of Pf Ag-specific IL-10(+)CD4(+) T cells in cord blood mononuclear cell cultures from these neonates. Generation and maintenance of IL-10(+) T cells in utero may thus contribute to suppression of APC function and Pf Ag-induced Th1 responses in newborns born to mothers with placental malaria at delivery, which may increase susceptibility to infection later in life.

Animals↗
PubMed2005

Neonatal and maternal immunological responses to conserved epitopes within the DBL-gamma3 chondroitin sulfate A-binding domain of Plasmodium falciparum erythrocyte membrane protein 1.

Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) mediates the adherence of P. falciparum-infected erythrocytes to placental syncytiotrophoblasts via interactions with chondroitin sulfate A (CSA), a characteristic of pregnancy-associated malaria. Pregnancy-associated malaria predicts increased susceptibility of newborns to malaria, and it is postulated that transplacental passage of parasite antigen induces immune regulatory activity in the neonate. We wished to examine the immune responsiveness to a CSA-binding domain of PfEMP1, the DBL-gamma3 domain, in cord and maternal venous blood obtained from pregnancies with various histories of P. falciparum infection. We assessed in vitro T-cell cytokine and plasma immunoglobulin G (IgG) and IgM responses to four peptides corresponding to highly conserved regions of a DBL-gamma3 domain common to central African parasite isolates. The presence of placental P. falciparum infection at delivery was associated with elevated frequencies of DBL-gamma3 peptide-specific CD3+ interleukin-10-positive T cells in cord blood, while treatment and clearance of infection prior to delivery was associated with elevated frequencies of CD3+ gamma interferon-positive T cells. DBL-gamma3 peptide-specific IgM antibodies were detected in 12 of 60 (20%) cord plasma samples from those born to mothers with P. falciparum infection during pregnancy. Consistent with polyclonal anti-PfEMP1 antibody responses that are associated with protection against pregnancy-associated malaria, the presence of maternal IgG antibodies with specificity for one of the DBL-gamma3 peptides showed a parity-dependent profile. These data demonstrate that peptides corresponding to conserved regions of the DBL-gamma3 domain of PfEMP1 are immunogenic in P. falciparum-infected mothers and their offspring.

Amino Acid Sequence↗
PubMed2002

Freeze-drying using vacuum-induced surface freezing.

A method of freezing during freeze-drying, which avoids undercooling of a solution and allows growth of large, dendritic ice crystals, was investigated. Aqueous solutions of mannitol, sucrose, or glycine were placed under a chamber vacuum of approximately 1 mbar at a shelf temperature of +10 degrees C. Under these conditions, the solutions exhibit surface freezing to form an ice layer of approximately 1-3 mm thickness. On releasing the vacuum and lowering the shelf temperature to below the freezing point of the ice in the solution, crystal growth occurs to yield large, chimney-like ice crystals. The duration of primary drying of a frozen cake--as measured by using inverse comparative pressure measurement--was up to 20% shorter than when using a "moderate" freezing procedure (2 K shelf temperature per min). With mannitol, however, the residual moisture content of the final dried product was higher than with moderate freezing, and with sucrose and glycine there was no difference. These findings are related to the structures of the dried cakes formed during freezing, as examined by light microscopy and wide-angle X-ray diffraction. The introduction of an annealing step (4 h at a shelf temperature slightly above the onset melting point of the ice in the frozen cake) combined with the vacuum-induced surface freezing procedure maintains the rapid primary drying and produces a low residual moisture (0.2%) for the freeze-dried mannitol solution.

Crystallization↗
PubMed2002

Successful treatment of idiopathic febrile panniculitis (Weber-Christian disease) with thalidomide in a patient having failed multiple other medical therapies.

The authors describe a case of idiopathic nodular panniculitis (Weber-Christian disease) with recurrent febrile episodes resistant to glucocorticosteroids and methotrexate (MTX) in various combinations with hydroxychloroquine, azathioprine, cyclosporine, colchicine, and doxycycline. Thalidomide at 100 mg/day has induced a remission for 3 years.

Journal Article↗
PubMedDate not supplied

Ultrasonographic and radiographic assessment of uncomplicated secondary fracture healing of long bones in dogs and cats.

OBJECTIVES: To evaluate the use of ultrasonography (US) to detect bone healing in uncomplicated diaphyseal fractures of dogs and cats, and to compare these observations with detection of healing by radiography (RG). STUDY DESIGN: Clinical study. ANIMALS: Dogs (33) and cats (11). METHODS: RG and brightness mode US were used to follow uncomplicated secondary fracture healing. Fractures were examined at admission and then every 2-4 weeks until healed or implant removal. Temporal differences in definitive detection of healed fracture by imaging technique were examined by species, patient age, bone, and fracture type. RESULTS: US images obtained during uncomplicated secondary fracture healing were consistent with images of fracture healing described in humans. Mean time to US diagnosis of a healed fracture (mean 46 days) was significantly shorter than by RG (mean 66 days). Mean time until diagnosis of a healed fracture (US and RG) did not differ significantly between open and closed treatment. Patients </=7 months (n=9) healed significantly faster (P<.05) than animals aged 7-36 months (n=24) and animals >36 months (n=11), but there was no significant difference between the latter 2 groups. Diagnosis of a healed simple fracture by US was significantly quicker than for a comminuted fracture (P<.05), but no difference was noted when using RG. CONCLUSIONS: US can be used to evaluate secondary fracture healing in biologically treated fractures in dogs and cats. US permits detection of a healed fracture earlier than RG. CLINICAL RELEVANCE: Earlier diagnosis of a healed fracture by US can prevent unnecessarily long limb immobilization and allow earlier dynamization.

Animals↗
PubMedDate not supplied

Intra- and interobserver variability of ultrasonographic measurements of the thyroid gland in healthy Beagles.

The repeatability of ultrasonographic measurements of the canine thyroid gland was evaluated. The variability of three different parameters (the maximal length, width, and height) within observer, between observer and between dogs was assessed based on three different measurements made by each of three observers infive healthy beagle dogs. From the three parameters, the volume of the gland was estimated using a formula of a rotation ellipse. The height and the volume had the lowest intra- and interobserver variability, while measurements of the length had the biggest intra- and interobserver variability. The mean values, with their 90% confidence interval were: height = 0.53cm [0.33-0.73], length = 2.45cm [2.04-2.85], width = 0.62cm [0.46- 0.78], volume = 0.38 cm3 [0.20-0.55].

Animals↗
PubMedDate not supplied

Comparison of the ultrasonographic appearance of osteochondrosislesions in the canine shoulder with radiography, arthrography, and arthroscopy.

Osteochondrosis lesions in 29 shoulder joints (from 20 dogs) were evaluated with ultrasound (US) and the results were compared with survey radiography, arthrography, and arthroscopy. US was performed with a 7-12 MHz linear matrix transducer which was placed in cranio-caudal direction just distally to the acromion while the joint was adducted and maximally endorotated to visualize the caudal aspect of the humeral head. With US, the subchondral defect was completely visible in 21 joints and partially visible in 8 joints. The length of the subchondral defect measured on US was comparable with the length measured on survey radiographs. In two joints, the cartilage flap was mineralized and thus already visible on survey radiographs. The mineralized flap was visible on US as a straight hyperechoic line above the subchondral defect. In the other joints, survey radiographs could not assess the status of the articular cartilage. In 17 joints, the presence of a cartilage flap or cartilage fissuring was suspected based on the presence of a second hyperechoic line at the base of the subchondral defect, and this suspicion was confirmed by arthroscopic examination in 16 joints and also by arthrographic examination in 15 joints. One joint that was suspected of having a cartilage flap on US was normal on arthroscopy and arthrography. When US revealed only focal thickening of the anechoic cartilage layer (5 joints), the joints appeared normal on arthroscopic and arthrographic examination. Of the four joints where the subchondral defect was irregular and covered by heteroechogeneous material on US, arthroseopy revealed the presence of a lesion resembling chondromalacia in two joints, the presence of a small cartilage flap in one joint and the presence of scar tissue underneath the flap at the level of the subchondral defect in one joint. In conclusion, US is a helpful imaging modality in the identification of osteochondritic lesions in the canine shoulder joint. US also appears to be a satisfactory imaging tool for identifying lesions such as joint mice, joint effusion, and distinct new bone formation.

Animals↗
PubMedDate not supplied

Power Doppler assessment of the neovascularization during uncomplicated fracture healing of long bones in dogs and cats.

The aims of this prospective study were to test the feasibility of assessing neovascularization with power Doppler ultrasonography and to investigate its usefulness to follow fracture healing of long bones in dogs and cats. A total of 51 patients (44 dogs and seven cats) were followed. Fracture types differed from simple to comminuted. Therapy ranged from external coaptation to plate osteosynthesis. Patients were followed with radiography, B-mode real time and power Doppler ultrasonography every 2-4 weeks until the fracture was healed. All fractures healed uneventfully. A semi-quantitative numerical score based on signal intensity, vessel area, and number of Doppler signals was assigned and the mean value was used to compare patients and examinations. Time postoperatively was divided into periods of 10 days. No Doppler signal was present during the first 10 days. The mean of the scores was highest between 11 and 20 days postoperatively and the median of the scores peaked between 21 and 30 days. A gradual decrease was seen thereafter. The mean of the scores was zero at 71-80 days and the median at 51-0 days postoperatively for the grouped results. In all positive power Doppler examinations, signals were present in and close to the callus. In seven patients (five dogs and two cats) signals were also present in the peripheral soft tissues in one of the follow up examinations. The normal healing process of fractured bones can be visualized using power Doppler ultrasonography and follows a distinctive time-dependent pattern.

Animals↗