Cell-based bone reconstruction therapies-principles of clinical approaches

Cell-based bone tissue engineering is a rapidly evolving therapy option in bone reconstruction strategies. Some cell-driven approaches, especially the biophysical stimulation of the host cell population surrounded by the bone defect, are common treatment methods in maxillofacial surgery. Others, such as autologous cell implantation, have now gained acceptance for clinical trials. More advanced or complex therapeutical options (extracorporeal tissue engineering, stem cell use, genetic engineering) have been tested in preclinical investigations but have not reached the level of clinical use. Two different aspects are of special relevance in cell-based bone reconstruction therapies. The source of cells used to regenerate bone (discussed in detail in a complementary review in this issue of The International Journal of Oral and Maxillofacial Implants) as well as the principal approach of a cell-driven bone regeneration therapy influence the outcome of such engineering strategies. All of the cell-driven repair strategies are under intensive investigation in an effort to provide surgeons with a limitless supply of tissue for bone repair and reconstruction in future procedures. An overview of the basic biological aspects as well as the inherent constraints of different cell-based approaches are given in this paper.     

Increase of the M phenotype among erythromycin-resistant Streptococcus pneumoniae isolates from Spain related to the serotype 14 variant of the Spain9V-3 clone

Between 1998 and 2003 the rate of erythromycin resistance among pneumococci in Spain was 34.4%. Although the MLS(B) phenotype was prevalent (94.7%), the rate of the M phenotype increased from 3.3% to 8.9% (P < 0.01). Clonal dissemination of mef(E)-carrying strains of serotype 14 variant of the Spain(9V)-3 clone was the major contributor to this increase.     

Human red blood cell Wright antigens: a genetic and evolutionary perspective on glycophorin A-band 3 interaction

The Wright (Wra/Wrb) blood group polymorphism is defined by an allelic change (Lys658Glu) in the band 3 protein; nevertheless, the Wrb antigen apparently requires glycophorin A (GPA) for surface presentation. To gain insight into the structural basis for this protein-protein interaction and delineate its relationship with Wrb antigen expression, we investigated GPA and band 3 sequence polymorphisms occurring in rare humans and nonhuman primates. The lack of GPA or amino acid residues 59 through 71 of GPA results in the absence of Wrb from human red blood cells (RBCs) exhibiting the MkMk, En(a-), or MiV phenotype. However, the SAT homozygous cells carried a Glu658 form of band 3 and a hybrid glycophorin with the entire GPA extramembrane domain from residues 1 through 71, yet expressed no Wrb antigen. This finding suggests that formation of the Wrb antigenic structure is dependent on protein folding and that the transmembrane junction of GPA is important in maintaining the required conformation. Comparative analyses of GPA and band 3 homologues led to the identification in the interacting regions of conserved and dispensable amino acid residues that correlated with the Wrb positive or negative status on nonhuman primates. In particular, the chimpanzee RBCs cells expressed Wrb and the Glu658 form of band 3, which is identical to humans, but their GPA contained the Gly rather than Arg residue at position 61. Taken together, the results suggest that (1) Arg61 of GPA and the proposed Arg61-Glu658 charge pair are not crucial for Wrb antigen exhibition and (2) the role of GPA for interaction with band 3, including Glu658, probably involves a number of amino acid residues located in the alpha-helical region and transmembrane junction.     

B cell growth factor-induced proliferation of hairy cell lymphocytes and inhibition by type I interferon in vitro

Malignant B cells from hairy cell leukemia (HCL) patients are unable to proliferate when stimulated with standard B cell mitogens. Using chromatographically purified B cell growth factor (BCGF), HCL can be stimulated to proliferate as assessed by incorporation of tritiated thymidine [3HTdR] into DNA. Proliferation was found to be time dependent, with no detectable 3H-TdR incorporation in up to three days of culture, and significant stimulation evident at days 6 and 10. The presence of 10% BCGF in culture was an absolute requirement for HCL proliferation; however, this BCGF-induced DNA synthesis could be further augmented by the addition of anti-immunoglobulin heavy chain antibodies. BCGF-induced proliferation was abrogated in six of six patients by addition of 1,000 U/mL of recombinant alpha 2-interferon (IFN) at day 0, although 1,000 U/mL of recombinant gamma-IFN had no inhibitory effect in five of six patients studied. Specific cellular receptors for type I IFN were demonstrated in HCL by inhibition of binding of 125I-alpha 2-IFN by a 40-fold excess of unlabeled alpha 2 or beta IFN with no inhibition by unlabeled gamma-IFN. These data demonstrate that malignant HCL lymphoblasts express specific type I IFN receptors and that type I, but not type II IFN, can inhibit growth factor-induced DNA synthesis by hairy cells in vitro. They further suggest a direct antiproliferative mechanism of action for IFN in HCL and predict equivalent clinical activity by either alpha or beta, but not gamma IFN in this malignancy.     

Interrelationships of bile acid and phospholipid fatty acid species with cholesterol saturation of duodenal bile in health and gallstone disease.

The relative amount of cholesterol and the fatty acid composition of phosphatidylcholines in bile can be influenced by the bile acid species secreted. To search for a contribution of secondary bile acids and of phosphatidylcholines to supersaturation of bile in gallstone disease, we compared the relative amount of cholesterol and the biliary composition of bile acids and of phospholipid fatty acids in cholecystokinin-stimulated duodenal bile of 22 female gallstone patients and 16 healthy controls and analyzed the interrelationships of these bile constituents. Gallstone patients had higher molar percentages of cholesterol than did controls (10.2 +/- 3.2 vs. 6 +/- 1.5 mol%; p less than 0.001) and demonstrated a trend toward larger fractions of deoxycholic and lithocholic acids. By linear models, variation of cholesterol saturation could be predicted (p less than 0.001) up to 53% by the bile acid pattern and up to 81% by the fatty acid pattern of phospholipids. Linear path analysis (goodness-of-fit index = 0.973) confirmed the tight relationship between phospholipid fatty acids (positive: oleic, arachidonic; negative: linoleic, palmitoleic) and the relative amount of cholesterol; more than half the influence of cholic, deoxycholic and lithocholic acids on the relative amount of cholesterol could be explained indirectly by their influence on the phospholipid fatty acid pattern. We conclude that the relationships examined by path analysis support the working hypothesis that secondary bile acids contribute to supersaturation of bile mainly by changing the fatty acid pattern of the secreted phospholipids (presumably the pattern of phosphatidylcholines), which increases the molar ratio of cholesterol/phospholipids in bile.     

Epidemiology and molecular basis of penicillin-resistant Neisseria meningitidis in Spain: a 5-year history (1985-1989).

Penicillin-resistant (penr) clinical isolates of Neisseria meningitidis, which do not produce beta-lactamase, were first identified in Spain in 1985; the frequency of their recovery, which has been increasing in the past few years, reached 20% in 1989. Serogrouping, determination of serotypes and subtypes, and multilocus enzyme electrophoresis of the penr strains showed an extensive diversity. Resistance is due, at least in part, to a decreased affinity of penicillin-binding protein (PBP) 2 for penicillin. Similar low-affinity forms of PBP 2 are also found in penr isolates of Neisseria lactamica, Neisseria polysaccharea, and Neisseria gonorrhoeae. Genetic transformation of an N. meningitidis type strain to low-level penicillin resistance with DNA from resistant meningococci and other Neisseria species resulted in transformants that possessed low-affinity forms of PBP 2. These altered forms of PBP 2 have been shown to arise from recombinational events that replace parts of the PBP 2 gene with the corresponding regions from the PBP 2 genes of commensal Neisseria species.     

Management of ��floating elbow�� in children

Background:

Supracondylar fractures associated with ipsilateral forearm fractures, aptly termed as “floating elbow” is a rare injury in children after a fall from height. The various authors have reported their results with conservative treatment of one or both injuries to aggressive emergency operative fixation of both components.

Materials and Methods:

During a period of three years, the author managed four cases of floating elbow in children. All cases were managed by closed reduction and pinning of both components of the injury.

Results:

All patients recovered full elbow range of motion at three months followup and were rated as excellent as per modified Flynn''s criteria. None of the patients developed cubitus varus deformity, complications related to the pins or delayed union.

Conclusions:

Early closed reduction and K wire fixation of both components of this injury gives better stability and prevents development of complications like compartment syndrome and elbow deformities.     

Neck neoplasms: MR imaging. Part I. Initial evaluation

Untreated neoplasms of the neck (tumors of the oropharynx, supraglottic area, carotid body, and thyroid, in addition to malignant lymphadenopathy) were evaluated in 23 patients with magnetic resonance (MR) imaging. The results were compared with computed tomographic (CT) scans in 20 patients. Contrast between tumor and fat was best on relatively T1-weighted images (500/30-35 [TR msec/TE msec]), whereas separation of tumor and muscle was best with relatively T2-weighted pulse sequences (1,500/90). Balanced images (1,500/30-35) provided best overall image quality and best demonstrated vascular anatomy. MR imaging was usually superior to CT in showing the relationship of tumor mass to muscle. MR imaging and contrast material-enhanced CT were equivalent in most patients in defining vascular anatomy, but MR imaging was superior when intravenous contrast material was not administered. However, CT was more helpful in showing bone and cartilage anatomy, and in some patients CT also was better in showing airway abnormalities. Despite these limitations, MR imaging is a promising imaging technique for studying neoplasms of the neck.     

Improvement of wound tissue repair by chitosan films containing (–)‐borneol,a bicyclic monoterpene alcohol,in rats

The aim of this study was to investigate the wound‐healing activity of (–)‐borneol (BOR) incorporated in chitosan film on healing protocol in rodents. To assess the BOR wound‐healing potential, male Wistar rats were subjected to a full‐thickness excisional wound. The animals were divided into three groups: dressed with chitosan‐based film (QUIN); dressed with chitosan‐based film containing 0·5% BOR (QUIBO05); or dressed with chitosan‐based film containing 1% BOR (QUIBO1). Dressing the wound areas and histological analysis were performed on the 3rd, 7th, 14th, and 21st days. The myeloperoxidase (MPO) activity was assessed on the third and seventh days after surgical procedures. Wounds dressed with chitosan‐based film containing BOR reduced significantly the MPO activity (P < 0·001), showed significantly larger wound retraction rates (7 days, P < 0·05), improved the granulation reaction, and also provided better collagenisation density and arrangement during wound healing. It is suggested that BOR modulates the wound‐healing process and is a promising compound to be used in wound care. This product may be quite useful in improving wound healing and could be a new biotechnological product with healing properties and clinical application. Further ongoing studies will enable us to understand the precise mechanisms whereby BOR improves the wound‐healing process.