Dimethyl sulfoxide (DMSO) is the cryoprotectant of choice for most animal cell systems since the early history of cryopreservation. It has been used for decades in many thousands of cell transplants. These treatments would not have taken place without suitable sources of DMSO that enabled stable and safe storage of bone marrow and blood cells until needed for transfusion. Nevertheless, its effects on cell biology and apparent toxicity in patients have been an ongoing topic of debate, driving the search for less cytotoxic cryoprotectants. This review seeks to place the toxicity of DMSO in context of its effectiveness. It will also consider means of reducing its toxic effects, the alternatives to its use and their readiness for active use in clinical settings.

Surgical experience with 2959 allogeneic and xenogeneic dense connective tissue grafts (1767 of fascia lata, 909 of pericardium, and 283 of dura mater), used in 2665 neurosurgical operations performed in the course of 20 years (1976 to 1995) is reported. Duraplasty using either allogeneic or xenogeneic grafts has had a similar, and favourable clinical outcome. Nevertheless, the pliable deep frozen fascia lata grafts, which could be used in any location, have been reserved for sella turcica plugging, anterior cranial base plasty, aneurysmal wrapping, and surgery of lipomyelomeningocele. Pericardium and dura mater grafts were in the majority of cases used over the brain convexity and posterior cranial fossa. Ovine pericardium proved to be superior to bovine and allogeneic pericardia because of its workability, flexibility, reduced thickness, and better transparency. Postsurgical complications occurred in 7.3%, and they were: 1) cerebrospinal fluid fistulas in 2.8%; 2) meningites in 2.3% (aseptic 1.4%, bacterial 0.8%, and tumoural 0.1% meningites); 3) pseudomeningoceles in 2.2%; 4) wound infections in 0.6%; 5) malresorptive hydrocephalus in 0.5%; and 6) adhesions to nerve tissue in 0.5%. The majority of complications healed without surgery. Forty-eight grafts (1.6%) failed to fulfil the requirements of the surgeon, and 46 of them were re-operated upon. Though another thirty-nine grafts healed successfully, 39 shunts (1.5%) had to be performed for malresorptive hydrocephalus (0.9%), and/or for a big pseudomeningocele (0.6%). So, the pure complication rate in 2665 duraplasties was 3.1%. The complex evaluation of the allogeneic and xenogeneic grafts (fascia, pericardium, and dura mater), used for duraplasty in neurosurgery during the last 20 years proved them, as remarkably good, with a success rates of 96.9%.

A 5-year experience with the glutaraldehyde-stabilized freeze-dried radiation-sterilized calf pericardium used as a dural substitute is reported. The structure of pericardium xenograft is compared with other collagenous materials used for duraplasty (allogeneic fascia lata and dura mater) by light and electron microscopy. The special neurosurgical techniques involved in using pericardium xenografts in the reconstruction of suboccipital dura mater in children are presented in detail.

The authors report on their 2 1/2-year clinical experience using a dural substitute, ovine pericardium, stabilized with 0.3% glutaraldehyde, flat freeze-dried, and sterilized with gamma-irradiation. Packaging of the ovine pericardium in double-plastic transparent bags allows simple storage in operating rooms and the opportunity for the surgeon to choose an ideal graft according to its shape, size, and plasticity. The ovine pericardia were examined histologically and by transmission and scanning electron microscopy in their native, freeze-dried, and irradiated forms. The final product is composed solely of pericardium fibrosum interwoven with artificially formed extracellular microcavities that serve as natural pores for the ingrowth of host tissue. The surfaces of the grafts appear smooth on the inner and rough on the outer sides. Sixteen grafts were examined macroscopically at autopsy and four of these were also examined histologically to illustrate their successful healing with no adhesion to the underlying brain. As a new dural substitute, ovine pericardium proved to be superior to bovine and allogeneic pericardia because of its workability, flexibility, and reduced thickness. In a study of 120 grafts, all but one healed without complications.

A systematic study, performed from 2017–2020 looked at the rate of positive post-pasteurization B. cereus findings, the quantity of B. cereus in pasteurized banked human milk (PBM), and the rate of B. cereus toxicogenic isolates from PBM. During the study period, 6815.71 L (30,943 tested bottles) of PBM were tested, with an average amount per year of 1703.93 L (7736 tested bottles). The PBM discard rate per year due to bacterial contamination varied between 8.7–10.0% and contamination with B. cereus was the most frequent reason. The total number of B. cereus positive tests was 2739 and the proportion of its positivity from all positive tests was between 56.7–66.6%. The prevalence of B. cereus positive tests rose significantly in the summer months. The production of enterotoxin was found in 3 of the 20 tested samples (15.0%). The B. cereus CFU-quantities in the PBM were below 10 CFU/mL in 80% of cases (16 of 20 samples tested). The quantitative data can be used in the risk assessment of cold storage of PBM at temperatures above zero and manipulation of PBM prior to its administration.

A series of 314 posterior fossa duraplasties in children were performed at the Department of Neurosurgery, Hradec Králové, over the past 33 years. Computed tomographic (CT) imaging was used to compare the healing of various kinds of collagenic dural substitutes--allogeneic fascia lata, allogeneic dura mater and xenogeneic pericardium. Early (8 days to 3 months following surgery) and late (1-18 years following surgery) axial CT scans with sagittal reconstruction for duraplasty in 55 children were evaluated. In early postoperative CT scans, epidural collections of cerebrospinal fluid, sanguinolent liquid or haematoma and/or pseudocysts or pseudomeningoceles appeared. In late CT follow-up, calcifications and ossifications in the "suboccipital coverings complex" (SCC) gradually developed and pseudocysts or pseudomeningoceles rarely persisted. It is concluded, on the basis of perioperative, clinical, and CT examinations, that posterior fossa duraplasties in children formed an important anatomical barrier and regardless of the type of graft had a favourable outcome; CT was shown to be a suitable method for the demonstration of dural grafts at the site of craniectomies.

Background. Vascular allotransplantations are performed worldwide in selected patients suffering from vascular prosthesis infection or critical limb ischemia. Either fresh or cryopreserved vascular allograft may be used. Objectives. In various points, we address several aspects (allograft procurement, cryopreservation and transplantation technique) of the program of vascular allotransplantations in the Czech Republic. Materials and methods. Vascular grafts retrieval has been done within multiorgan harvests using no-touch technique. Very short time of cold ischemia is achieved due to close cooperation with Tissue Establishment where the following processing of cryopreservation is performed. Meeting all necessary quality criteria is a prerequisity for releasing grafts for clinical application. Standardized thawing protocol and surgical handling aims to minimize microfractures before implantation. Results. Based on experimental and clinical work, the first validation of cryopreserved arterial and venous grafts for clinical use was performed between 2011 and 2013 in the Czech Republic. The developement of storage of vascular tissue in banks was stimulated in 2000-2010 by the issue of EU directives and national harmonized norms, aimed at assurance of high quality and safety of cells and tissues used for transplantations in humans. Conclusions. There are several crucial moments affecting final quality, including graft retrieval within a multiorgan harvest, short ischemic time, cryopreservation, and thawing technique used. The recommended surgical handling during implantation may also affect results and graft-related complications.

BackgroundThe aim of our experimental work was to assess morphological changes of arterial wall that arise during different thawing protocols of a cryopreserved human aortic root allograft (CHARA) arterial wall.MethodsThe experiment was performed on CHARAs. Two thawing protocols were tested: 1, CHARAs were thawed at a room temperature at +23°C; 2, CHARAs were placed directly into a water bath at +37°C.Microscopic samples preparationAfter fixation, all samples were washed in distilled water for 5 min, and dehydrated in a graded ethanol series (70, 85, 95, and 100%) for 5 min at each level. The tissue samples were then immersed in 100% hexamethyldisilazane for 10 minutes and air dried in an exhaust hood at room temperature. Processed samples were mounted on stainless steel stubs, coated with gold.ResultsThawing protocol 1: All 6 (100%) samples showed loss of the endothelium and damage to the subendothelial layers with randomly dispersed circular defects and micro-fractures without smooth muscle cells contractions in the tunica media.Thawing protocol 2: All 6 (100%) samples showed loss of endothelium from the luminal surface, longitudinal corrugations in the direction of blood flow caused by smooth muscle cells contractions in the tunica media with frequent fractures in the subendothelial layerConclusionAll the samples thawed at the room temperature showed smaller structural damage to the CHARA arterial wall with no smooth muscle cell contraction in tunica media when compared to the samples thawed in a water bath.