Biomedical subjects

I Olsen

Publications and source records attributed to I Olsen.

At least 19 recordsLinked to original sources

PubMed2006

Shedding of a soluble form of BMP receptor-IB controls bone cell responses to BMP.

Bone morphogenetic proteins (BMP) are members of the transforming growth factor beta (TGF-beta) superfamily and are involved in a wide variety of biological processes, including osteoblast differentiation and bone healing. The activities of the BMP are mediated by signal transduction via three BMP receptors (BMPR-IA, -IB and -II), which are thus essential for the biological actions of the BMP. Although the precise mechanisms which control the BMPR are not yet known, it is possible that post-translational regulation of these cell surface antigens by shedding could modulate their expression and thereby at least partly determine the response of the cells to the BMP. To test this possibility, the present study has examined whether soluble forms of the BMPR are produced by shedding from primary human bone cells in vitro. The results showed that human bone cells expressed both mRNA transcripts and antigens corresponding to BMPR-IA, -IB and -II. Incubation of the cells with phorbol 12-myristate 13-acetate (PMA), a potent inducer of proteolytic shedding, resulted in a pronounced decrease in cell surface expression of all three BMPR and, concurrently, the presence of "soluble" forms of these antigens in culture supernatants. Moreover, PMA treatment significantly reduced the level of BMP-2-induced Smad1/5 phosphorylation, a major early activation step in signal transduction initiated by BMP/BMPR interaction. It is notable that, while treatment of bone cells with interleukin-1beta (IL-1beta) also reduced the level of surface BMPR-IB, this inflammatory cytokine had no effect on BMPR-IA or -II levels, hence only the soluble form of BMPR-IB was detected. Furthermore, in addition to down-regulating BMP-2-induced Smad1/5 phosphorylation, IL-1beta also caused a reduction in the level of BMP-2-induced alkaline phosphatase activity and osteocalcin expression, both closely associated with bone cell differentiation. In conclusion, our study has provided evidence, for the first time, that BMPR can be modulated at the cell surface by the shedding of a soluble form of the antigen, resulting in a markedly diminished response to BMP-2 in vitro.

Blotting, Western↗

PubMed2006

Bone morphogenetic protein receptors and bone morphogenetic protein signaling are controlled by tumor necrosis factor-alpha in human bone cells.

Bone morphogenetic proteins (BMP) stimulate osteoblast differentiation by signal transduction via three BMP receptors (BMPR-IA, -IB and -II), whereas the inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) has been shown to suppress osteoblast differentiation. Although the mechanisms which regulate the BMPR are not yet known, it is possible that they may be negatively controlled by TNF-alpha, thereby inhibiting BMP-induced osteoblast differentiation. To test this hypothesis, we have examined the effects of TNF-alpha on BMPR-IA, -IB and -II expression and the functional consequences of this cytokine on BMPR-mediated functions in human bone cells. The results showed that although TNF-alpha down-regulated BMPR-IA and -II transcripts, it increased the level of BMPR-IB mRNA via a MAPK-dependent pathway. In marked contrast, however, TNF-alpha nevertheless caused marked down-regulation of the expression of the BMPR-IB surface antigen specifically. Moreover, the cytokine-induced decrease in BMPR-IB expression was found to be associated with the concurrent presence of a 'soluble' form of this antigen in supernatants of TNF-alpha-treated cultures. Furthermore, the TNF-alpha-induced loss of BMPR-IB was found to ablate BMP-2-stimulated bone cell functions, including phosphorylation of Smad1/5/8, alkaline phosphatase activity and osteocalcin expression. In conclusion, our study has provided evidence, for the first time, that BMPR can be differentially modulated by TNF-alpha at both the post-transcriptional and post-translational levels, with the TNF-alpha-induced shedding of the BMPR-IB antigen associated with a significantly diminished response to BMP-2 in vitro.

Antigens↗

PubMed2006

Genotypic relatedness of yeasts in thrush and denture stomatitis.

BACKGROUND/AIM: Candida is an opportunistic pathogen. Understanding its genetic characters might increase our understanding of the pathogenesis of candidosis. We examined the genetic relationships of yeasts from the most common forms of oral candidosis: thrush and denture stomatitis. METHODS: Yeasts were sampled from palate, buccal mucosa, gingival sulci/periodontal pockets and/or denture fitting surface of 19 thrush patients and 22 denture stomatitis patients. Random amplified polymorphic DNA and the Dendron computer-assisted program were used to determine the genotypic relatedness of the yeasts. RESULTS: A dendrogram generated from 105 thrush isolates had similarity coefficients (S(AB)) ranging from 0.58 to 1 with four clusters derived at S(AB) 68%. Another dendrogram was generated from 91 isolates from denture stomatitis, with S(AB) ranging from 0.59 to 1. Three clusters were established at S(AB) 71%. In a composite dendrogram incorporating the thrush and denture stomatitis data and orally healthy data compiled from a previous study, five genotypic clusters were generated at S(AB) 68%. Cluster II, the most dominant, comprised isolates from thrush, denture stomatitis and healthy conditions, while clusters III and IV contained yeasts mainly from thrush. CONCLUSIONS: Palatal yeast carriage was significantly increased in thrush and denture stomatitis, also after radiation, chemotherapy and denture wearing. The buccal mucosa was favorable for yeast colonization regardless of oral condition. Yeasts in thrush were more diverse than in conditions of oral health. The common clone (II) of infecting yeasts and commensals suggested that commensals could induce thrush and denture stomatitis, whereas the unique clones in thrush (III, IV) might have been established through strain replacement or maintenance with minor genetic variation.

Adolescent↗

PubMed2005

Effects of ultrasound on transforming growth factor-beta genes in bone cells.

Therapeutic ultrasound (US) is a widely used form of biophysical stimulation that is increasingly applied to promote fracture healing. Transforming growth factor-beta (TGF-beta), which is encoded by three related but different genes, is known to play a major part in bone growth and repair. However, the effects of US on the expression of the TGF-beta genes and the physical acoustic mechanisms involved in initiating changes in gene expression in vitro, are not yet known. The present study demonstrates that US had a differential effect on these TGF-beta isoforms in a human osteoblast cell line, with the highest dose eliciting the most pronounced up-regulation of both TGF-beta1 and TGF-beta3 at 1 hour after treatment and thereafter declining. In contrast, US had no effect on TGF-beta2 expression. Fluid streaming rather than thermal effects or cavitation was found to be the most likely explanation for the gene responses observed in vitro.

Acoustics↗

PubMed2005

DNA-DNA hybridization demonstrates multiple bacteria in osteoradionecrosis.

Bone necrosis secondary to radiation was previously attributed to trauma of devitalized bone and microbiological sepsis. However, conventional microbiological technique has failed to demonstrate microorganisms throughout osteoradionecrotic bone, claimed to be hypoxic, hypovascular and hypocellular. The aim of the present study was to examine such bone for bacteria using DNA-DNA hybridization. Compared to standard culture methods this technique enables the investigation of a vast number of bacteria in a fairly short time. Twelve deep medullary specimens from resected radionecrotic mandibles were studied. A multitude of bacterial species were detected, most of them anaerobic. Porphyromonas gingivalis was the most predominant organism, followed by Fusobacterium nucleatum subspecies polymorphum. All samples contained Actinomyces, Prevotella and F. nucleatum. The results of this study indicate that bacteria, particularly anaerobes, may play a more fundamental role in the pathophysiology of osteoradionecrosis than being merely surface contaminants.

Actinomyces↗

PubMed2005

Scanning and transmission electron microscopy demonstrates bacteria in osteoradionecrosis.

According to the current concept of osteoradionecrosis such tissue has become hypoxic, hypovascular and hypocellular and it has previously failed to show any bacterial growth. By using a molecular method, i.e., DNA-DNA hybridization, the presence of microorganisms in radionecrotic bone has formerly been demonstrated by us. The aim of the present study was to visualise bacteria in such bone by using scanning and transmission electron microscopy. Fourteen deep medullar specimens from resected radionecrotic mandibles were studied. Microorganisms were detected in 9 of the 14 samples (64%). The bacteria seen included rods, spirochetes and cocci, with rods being the predominant cells. One sample also contained yeasts (blastoconida and pseudohyphae). The data from this study strongly suggest an association between microorganisms and osteoradionecrosis. This may call for a revision of its presumed pathophysiology, including the possibility of an infectious etiology.

Adult↗

PubMed2005

Acute glomerulonephritis associated with streptococcus pyogenes with concomitant spread of streptococcus constellatus in four rural families.

We studied history, renal histopathology and microbiology of an epidemic of acute glomerulonephritis associated with throat infections and uncommon culture results in four neighbour families. A 40-year-old man (index patient) was referred to a university hospital for dialysis and kidney biopsy due to a suspected acute glomerulonephritis. An acute tonsillitis had preceded the condition. Penicillin treatment had been started four days before the discovery of renal failure. Throat swabs were positive for beta-hemolytic streptococci, group C (GCS). GCS were also found in throat cultures from his wife and two of their children. The bacteria were typed as Streptococcus constellatus. A third child had S. constellatus expressing Lancefield antigen group G. A neighbour and two of his children fell ill the following week with renal involvement. Throat swabs from both these children were positive for S. constellatus. His third child had erythema multiforme and S. constellatus in the throat while a fourth child had beta-hemolytic streptococci group A; Streptococcus pyogenes. Kidney biopsies on the index patient and his neighbour showed an acute diffuse prolipherative glomerulonephritis compatible with acute post-streptococcal nephritis and microbiological analysis of renal tissue revealed in both cases S. pyogenes and S. constellatus. The families had had much contact and had consumed unpasteurized milk from our index patient's farm. In four of seven persons in two additional neighbouring families S. constellatus was found in throat swabs during the same month while two persons carried Streptococcus anginosus expressing the Lancefield C antigen. In conclusion spread of S. constellatus coincided with the occurrence of four cases of acute glomerulonephritis. The two biopsied patients had both S. pyogenes and S. constellatus present in renal tissue. The epidemic either suggested that the outbreak of glomerulonephritis was due to S. pyogenes but coincided with the transmission and colonization of S. constellatus or that the S. constellatus strains were highly pathogenic or nephritogenic and that this organism can be transmitted in such cases.

Acute Disease↗

PubMed2004

Human alveolar bone cell adhesion and growth on ion-implanted titanium.

Surface characteristics play a vital role in determining the biocompatibility of materials used as bone implants. Calcium ion implantation of titanium was previously reported to enhance osseointegration and bone formation in vivo, although the lack of consistent and reproducible effects highlight the need to understand the basic mechanisms involved in the response of target cells to such surfaces. The aim of this study was therefore to measure the precise effects of ion implantation of titanium on bone cells in vitro. Alveolar bone cells were seeded on the surface of polished titanium disks implanted with calcium, potassium, and argon ions. Using radioisotopically tagged bone cells, the results showed that although the calcium ion implanted surface reduced cell adhesion, it nevertheless significantly enhanced cell spreading and subsequent cell growth. In contrast, few differences in bone cell behavior were observed between the potassium- and argon-implanted titanium and the control nonimplanted titanium disks. These findings suggest the possibility that the calcium-implanted surface may significantly affect the biocompatibility of titanium implants by enhancing bone cell growth. Surface modification by ion implantation could thus prove to be a valuable tool for improving the clinical efficacy of titanium for bone repair and regeneration in vivo.

Argon↗

PubMed2004

Gene expression profiling of bone cells on smooth and rough titanium surfaces.

Titanium (Ti) and Ti alloys are widely used as dental and orthopaedic implants, but the effects of the surface characteristics of these materials on the response of cells and target tissues is not well understood. The present study has therefore examined the effects of a rough Ti (RT) and a smooth Ti (ST) surface on human bone cells in vitro. Scanning electron microscopy showed attachment and spreading of cells on both surfaces. Expression profiling using ATLAS gene arrays showed marked differences in gene responses after 3 h of culture. A number of osteoblast genes were identified as "roughness response" genes on the basis of changes in expression on the RT compared with the ST surfaces. The surface roughness of Ti was thus found to have a profound effect on the profile of genes expressed by the bone cells, and suggests that improvements in the biological activity and possibly the clinical efficacy of these materials could be achieved by selective regulation of gene expression mediated by controlled modification of Ti surface.

Adult↗

PubMed2004

Processing, characterisation and biocompatibility of iron-phosphate glass fibres for tissue engineering.

Iron-phosphate glass fibres based on the CaO-Na2O-Fe2O3-P2O5 system have been processed and characterised via thermal, XRPD, dissolution rates, diameter and biocompatibility studies. The compositions investigated were fixed at 50mol% P2O5, and the CaO content was varied between 30, 35 and 40mol%. The Fe2O3 was added in low amounts from 1-5mol%, substituting it for the Na2O mol%. The number of Tc (crystallisation temperature) peaks detected from the thermal analysis traces only showed correlation with XRPD analysis, for five out of the 15 compositions investigated. It has been suggested that either the crystalline phases had very similar Tc temperatures or that the other phase(s) were present in very small quantities. There was a good match seen with number of Tm (melting temperature) peaks picked up from the DTA traces, with the number of phases identified from XRPD analysis. The main phases identified from XRPD were NaCa(PO3)3, CaP2O6 and NaFeP2O7. Using network connectivity (NC), predictions on Qn species present within the compositions investigated were made. The predicted species (metaphosphates) matched with phases identified from XRPD analysis. A decrease in dissolution rates for the bulk glass and glass fibres was seen with an increase in CaO mol%, along with an increase in Fe2O3 mol%. An increase in fibre dissolution rates was seen with a decrease in diameter size. The biocompatibility studies were conducted using a conditionally immortal muscle precursor cell line derived from the H-2Kb-tsA58 immortomouse. It was found that iron-phosphate glass fibres containing 4-5mol% Fe2O3 was sufficient for cell attachment and differentiation. It was seen that myotubes formed along the axis of the fibres (which was indicative of differentiation). The biocompatibility of these compositions was attributed to the enhanced chemical durability of the glass fibres.

Animals↗

PubMed2004

Roughness response genes in osteoblasts.

Titanium (Ti) and Ti alloys are widely used as dental and orthopedic implants, but the effects of the surface characteristics of these materials, including roughness, on the response of target tissues in vivo are not well understood. The present study has therefore examined the effects of a moderately rough (sand-blasted, acid-etched; SLA) Ti surface, a highly rough (plasma-sprayed; TPS) surface, and a smooth surface (SMO) on bone cells in vitro. X-ray photoelectron spectroscopy showed that these surfaces had similar surface chemistry, while scanning electron microscopy suggested that the SLA provided a transiently less biocompatible surface, with initially less well-attached cells. SLA also delayed bone cell growth compared with SMO, whereas the TPS surface elicited the greatest increase in cell numbers. In addition, expression profiling using the ATLAS gene array showed marked differences in gene responses after 3 h of incubation; this increased further after 24 h, with TPS generating the largest number of up- and down-regulated genes compared with SLA and SMO. A number of osteoblast genes were also identified as 'roughness' genes on the basis of their similar response on SLA and TPS, compared with SMO. These findings show, for the first time, that the surface roughness of Ti has a profound effect on the profile of genes expressed by bone cells and suggest that improvements in the biological activity and possibly the clinical efficacy of these materials could be achieved by selective regulation of gene expression mediated via modification of surface roughness.

Adult↗

PubMed2004

Phosphate glasses for tissue engineering: Part 1. Processing and characterisation of a ternary-based P2O5-CaO-Na2O glass system.

This paper presents the results of a study of the thermal properties, X-ray powder diffraction (XRPD), NMR and solubility of a range of phosphate-based glasses. Ion release and pH measurements were also obtained. The compositions were limited by fixing the P2O5 mol% content at 45, 50 and 55 mol%, and varying the CaO mol% at 30, 35 and 40 mol%. Initial data collected from thermal analysis showed an increase in the Tg values, with increasing CaO mol%, as expected. An increase in crystallisation temperatures with increasing CaO mol% was also observed, with sharp peaks indicating the presence of single and/or double phases. Initial XRPD data revealed that the 45 mol% P2O5 precipitated a calcium bridged trimetaphosphate (Na4Ca[PO3]6) at the sodium oxide contents of 25 and 20 mol%, but at the 15 mol% level two phases formed, NaCa[PO3]3 and Ca2P2O7. The 50 mol% P2O5 compositions showed two sodium phosphate-rich phases [NaCa(PO3)3 and NaPO3] for the lower calcium containing compositions, and a calcium phosphate phase [CaP2O6] for compositions with higher CaO mol%. The 55 mol% P2O5 compositions, exhibited similar phases for the lower calcium containing compositions, and a single calcium phosphate phase (CaP2O6) was identified for the higher calcium containing composition. NMR analysis revealed the presence of Q1 and Q2 species in the 45 mol% P2O5 compositions, and Q2 species present in the 50 and 55 mol% P2O5 compositions. The non-linear data obtained from the thermal and solubility analyses conducted were attributed to the packing density of the 45 mol% P2O5 compositions. The solubility was seen to decrease with increasing CaO mol%, for all the glasses investigated, and all the compositions showed a gradual decrease in pH with time, and this was accounted for by the release of Na+ and Ca2+ ions into solution.

Biocompatible Materials↗

PubMed2004

Phosphate glasses for tissue engineering: Part 2. Processing and characterisation of a ternary-based P2O5-CaO-Na2O glass fibre system.

This paper presents the results of a study of the thermal properties, solubility and dimensions of a range of phosphate-based glass fibres (PB-GFs). The glass compositions were limited by fixing the P2O5 content to 45, 50 and 55 mol%, and varying the CaO mol% at 30, 35 and 40. PB-GFs were obtained from the 50 and 55 mol% P2O5 compositions; however, we were unable to obtain fibres from the 45 mol% compositions. This was linked to the cross-linked density, network connectivity and average chain length of the compositions studied. With regards to thermal parameters investigated, initial data showed an increase of the Tg and crystallisation temperatures with increasing CaO mol% at each fixed phosphate content. A decrease in Tg temperatures was also observed with increasing P2O5 content to 55 mol%. The crystallisation temperatures obtained for compositions with fixed phosphate at 55 mol%, showed a reverse pattern, with a decrease in values as compared to the fixed 50 mol% phosphate compositions. The diameters of the fibres all decreased with increasing RPMs as expected, and the solubility also increased with increasing RPMs. This was related to the increased surface area of the higher RPM fibres. There was also a decrease seen in solubility with increasing CaO mol%.

Biocompatible Materials↗

PubMed2004

Beta-lactamase production and antimicrobial susceptibility of subgingival bacteria from refractory periodontitis.

This study assessed the extent of beta-lactamase-producing bacteria in subgingival plaque samples obtained from 25 patients with refractory marginal periodontitis in the USA. beta-Lactamase-positive isolates were characterized using commercial diagnostic kits and partial sequencing of the 16S rRNA gene. The susceptibilities to different antimicrobial agents were tested and, in addition, the isolates were screened for the presence of extended spectrum beta-lactamases (ESBLs). beta-lactamase-producing bacteria were detected in 18 (72%) patients. The most prominent beta-lactamase-producing organisms belonged to the anaerobic genus Prevotella. Other enzyme-producing anaerobic strains were Fusobacterium nucleatum, Propionibacterium acnes and Peptostreptococcus sp. Facultative bacteria, such as Burkholderia spp., Ralstonia pickettii, Capnocytophaga spp., Bacillus spp., Staphylococcus spp. and Neisseria sp., were also detected among the enzyme-producers. Minimum inhibitory concentrations (MICs) of ampicillin and amoxicillin were in the range 1.5-256 micrograms/ml and 4-256 micrograms/ml, respectively, for the isolates of the Prevotella species. All Prevotella isolates were susceptible to amoxicillin/clavulanate and metronidazole, but they showed variable resistance to tetracyclines. Two of the Prevotella isolates had high MICs of cefotaxime and ceftazidime. ESBL activity was not detected in any of the beta-lactamase-producing isolates by the Etest method. Thus, our study demonstrated a wide variety of beta-lactamase-producing bacteria that may play a role in refractory periodontal disease.

Adult↗

PubMed2004

Changes in transforming growth factor-beta1 in gingival crevicular fluid following periodontal surgery.

OBJECTIVES: Growth factors play a major part in wound healing, including in the periodontium. However, the presence of growth factors in gingival crevicular fluid (GCF) in humans during periodontal wound healing has not yet been determined. Our hypothesis is that such factors are present in GCF and that changes in their levels might be of value as a prognostic marker of wound-healing activity and therapeutic progress following periodontal surgery. The aim of this study was therefore to measure transforming growth factor-beta1 (TGF-beta1) in GCF collected from sites that have undergone guided tissue regeneration (GTR) and conventional flap (CF) surgery and to compare these with GCF collected from unaffected healthy sites. MATERIALS AND METHODS: GCF samples were collected, using filter paper strips, at baseline (pre-surgical) and then at intervals up to 26 weeks from 16 patients undergoing GTR and from 11 patients undergoing CF surgery. After elution and acid treatment, TGF-beta1 levels were measured by ELISA. RESULTS: Treatment of periodontal defect sites significantly reduced the mean probing pocket depth (PPD) and improved the mean lifetime cumulative attachment loss (LCAL). Average GCF volumes also significantly increased at all sites at 2 weeks post-surgery and thereafter declined to baseline levels, except at the GTR test sites that were still elevated at 7 weeks. TGF-beta1 could be detected in almost all GCF samples, and 2 weeks after surgery, the average levels increased two-fold at the surgically treated but not at the control sites, which remained unchanged. CONCLUSION: TGF-beta1 is readily detectable in GCF and increases transiently following periodontal surgery. This suggests that changes in the levels of this growth factor in GCF might be useful for monitoring the progress of periodontal repair and regeneration.

Adult↗

PubMed2003

Adhesion of bone cells to ion-implanted titanium.

The use of ion-implantation to encourage osseointegration has been investigated using an in vitro model cell culture system and surface analysis. Polished titanium discs were implanted with calcium, potassium and argon ions. The adhesion of bone-derived cells was measured using radioactively labeled cells and the morphology examined using scanning electron microscopy. Similar numbers of cells were found to adhere to the potassium and argon-implanted titanium as to control (non-implanted) titanium. However, adhesion to the calcium-implanted titanium discs was significantly reduced. Moreover, although the cells were found to be well spread on the calcium and potassium-implanted titanium, a much greater proportion of cells appeared to remain rounded and poorly attached on the argon-implanted surface. These differences are discussed in relation to the observed surface roughness and chemistry, which were assessed using interferometry and X-ray photoelectron spectroscopy, respectively.

Journal Article↗

PubMed2003

DNA injection in combination with electroporation: a novel method for vaccination of farmed ruminants.

Injection of plasmid DNA encoding antigens into rodents followed by electroporation improved the immune response when compared with injection without electroporation (Widera et al. J Immunol 2000;164:4635-40; Zucchelli et al. J Virol 2000;74:11598-607; Kadowaki et al. Vaccine 2000;18:2779-88). The present study describes the extension of this technology to farm animals, by injecting plasmid DNA encoding mycobacterial antigens (MPB70, Ag85B and Hsp65) into the muscles of goats and cattle using two different types of electrodes, both allowing DNA delivery at the site of electroporation. The animals were vaccinated under local anaesthesia without any observed immediate or long-term distress or discomfort, or any behavioural signs of muscle damage or pathological changes after the electroporation. DNA-injected and electroporated goats showed increased humoral response after the primary vaccination when compared with nonelectroporated animals. Improved T-cell responses following electroporation were observed in hsp65 DNA-vaccinated cattle. DNA injection with or without electroporation did not compromise the specificity of the tuberculin skin test. In conclusion, a protocol applying in vivo electroporation free of side effects to farmed ruminants was established. In addition, we show that DNA vaccination in combination with electroporation can improve the primary immune responses to the encoded antigens.

Animals↗

PubMed2002

The effect of MgO on the solubility behavior and cell proliferation in a quaternary soluble phosphate based glass system.

This paper presents a systematic study of the MgO-CaO-Na(2)O-P(2)O(5) glass system, which has great potential to be used as temporary hard and soft tissue implant materials. An overall study of solubility behavior of ternary and quaternary-based phosphate glass system have been carried out in order to understand the out-leaching progress of different ions and to determine their effect on cell proliferation. Originally, soluble phosphate based glasses within the ternary glass system of Na(2)O-CaO-P(2)O(5) have been developed to create a simple baseline system. This paper, however, presents the development of this system by introducing magnesium oxide as a partial calcium oxide substitute and solubility behaviors as well as cell studies have been carried out to check the effect on magnesium ions. Glasses have been prepared via standard glass melting techniques and their solubility behavior has been tested in distilled water via simple weight loss, pH and ion measurements. The way the glasses dissolve is an inverse exponential behavior which is mirrored by the calcium ion release. Other ions show a less exponential behavior. The MTT test has been used to check preliminary in vitro studies on a human MG63 cell line and the result indicates that cell proliferation is increased for glasses with minimal CaO substitution.

Journal Article↗