A review on diatom biosilicification and their adaptive ability to uptake other metals into their frustules for potential application in bone repair.

Diatoms are unicellular eukaryotic algae that have a distinctive siliceous cell wall (frustule) with unique architectures. The nanotopography of the frustule is perfectly replicated between generations, offering a source of highly intricate and identical silica microparticles. In recent years, the ability to alter their cell wall chemistry both in terms of functionalisation with organic moieties or by incorporation of the metal ions in their frustules has increased interest in their utility for catalysis technologies, and semiconductor and biomedical applications. Herein we review the fundamental biological mechanisms in which diatoms produce their frustule and their ability to substitute different metal ions in their frustule fabrication process. The review focuses on the potential of diatom frustules as a naturally derived biomaterial in bone tissue engineering applications and how their cell walls, comprising biogenic silica, could either partially or fully incorporate other bone therapeutic metal ions, e.g., titanium or calcium, into their frustule. The use of diatom frustules in bone repair also potentially offers a 'greener', more environmentally friendly, biomaterial as they can naturally synthesise oxides of silicon and other metals into their frustules under ambient conditions at a relatively neutral pH. This process would negate the use of harsh organic chemicals and high-temperature processing conditions, often used in the fabrication of silica based biomaterials, e.g., bioactive glass.

A comparison of the degradation behaviour of 3D printed PDLGA scaffolds incorporating bioglass or biosilica.

Silica incorporation into biomaterials, such as Bioglass and Si-substituted calcium phosphate ceramics has received significant attention in bone tissue engineering over the last few decades. This study aims to explore the dissolution behaviour of natural biosilica isolated from a freshwater diatom, Cyclotella meneghiniana, that has been incorporated into 3D printed poly (DL-lactide -co - glycolide) (PDLGA) scaffolds using extrusion and additive manufacturing. In the study, two different dry weight percentage (1 wt% & 5 wt%) of diatom-silica were incorporated into PDLGA scaffolds that were then degraded in phosphate buffered saline (PBS) cell free media. In addition, pure PDLGA scaffolds and 5 wt% Bioglass scaffolds were used as control groups. The degradation study was performed over 26-weeks. The release rate of Si4+ ions from diatom-PDLGA scaffolds was found to increase exponentially with respect to time. The compressive strength of scaffolds was also measured with the Diatom-PDLGA scaffolds found to maintain their strength for longer than either pure PDLGA scaffolds or 5 wt% Bioglass scaffolds. 13C NMR data showed that diatom biosilica containing scaffolds had less degradation than pure or bioglass-containing scaffolds at comparable time-points. Overall, the Diatom-PDLGA scaffolds were found to have more desirable physiochemical properties for bone repair compared to Bioglass.

Filament extrusion of bioresorbable PDLGA for additive manufacturing utilising diatom biosilica to inhibit process-induced thermal degradation.

Bone scaffolds are often fabricated by initially producing custom-made filaments by twin-screw extruder and subsequently fabricating into 3D scaffolds using fused deposition modelling. This study aims to directly compare the effect of two alternative silica-rich filler materials on the thermo-mechanical properties of such scaffolds after extrusion and printing. Poly (DL-lactide-co-glycolide) (PDLGA) was blended with either 45S5 Bioglass (5 wt %) or Biosilica (1 and 5 wt%) isolated from Cyclotella meneghiniana a freshwater diatom were tested. Diatom-PDLGA was found to have similar mechanical strength and ductility to pure-PDLGA, whereas Bioglass-PDLGA was found induce a more brittle behaviour. Bioglass-PDLGA was also found to have the lowest toughness in terms of energy absorption to failure. The TGA results suggested that significant thermal degradation in both the Bioglass filaments and scaffolds had occurred as a result of processing. However, diatom biosilica was found to inhibit thermal degradation of the PDLGA. Furthermore, evidence suggested the agglomeration of Bioglass particles occurred during processing the Bioglass-PDLGA filaments. Overall, diatom biosilica was found to be a promising candidate as a bone filler additive in 3D printed PDLGA scaffolds, whereas Bioglass caused some potentially detrimental effects on performance.

Enhanced release of calcium phosphate additives from bioresorbable orthopaedic devices using irradiation technology is non-beneficial in a rabbit model: An animal study.

OBJECTIVES: Bioresorbable orthopaedic devices with calcium phosphate (CaP) fillers are commercially available on the assumption that increased calcium (Ca) locally drives new bone formation, but the clinical benefits are unknown. Electron beam (EB) irradiation of polymer devices has been shown to enhance the release of Ca. The aims of this study were to: 1) establish the biological safety of EB surface-modified bioresorbable devices; 2) test the release kinetics of CaP from a polymer device; and 3) establish any subsequent beneficial effects on bone repair in vivo. METHODS: ActivaScrew Interference (Bioretec Ltd, Tampere, Finland) and poly(L-lactide-co-glycolide) (PLGA) orthopaedic screws containing 10 wt% ß-tricalcium phosphate (ß-TCP) underwent EB treatment. In vitro degradation over 36 weeks was investigated by recording mass loss, pH change, and Ca release. Implant performance was investigated in vivo over 36 weeks using a lapine femoral condyle model. Bone growth and osteoclast activity were assessed by histology and enzyme histochemistry. RESULTS: Calcium release doubled in the EB-treated group before returning to a level seen in untreated samples at 28 weeks. Extensive bone growth was observed around the perimeter of all implant types, along with limited osteoclastic activity. No statistically significant differences between comparative groups was identified. CONCLUSION: The higher than normal dose of EB used for surface modification did not adversely affect tissue response around implants in vivo. Surprisingly, incorporation of ß-TCP and the subsequent accelerated release of Ca had no significant effect on in vivo implant performance, calling into question the clinical evidence base for these commercially available devices.Cite this article: I. Palmer, S. A. Clarke, F. J Buchanan. Enhanced release of calcium phosphate additives from bioresorbable orthopaedic devices using irradiation technology is non-beneficial in a rabbit model: An animal study. Bone Joint Res 2019;8:266-274. DOI: 10.1302/2046-3758.86.BJR-2018-0224.R2.

Feedlot performance and immune function analysis of implanted and non-implanted steers selected for alcohol dehydrogenase 1 C (ADH1C) genotype and fed a low vitamin A diet.

Previous studies have shown that the interaction between limiting vitamin A (VA) and an alcohol dehydrogenase 1 C (ADH1C) variant in beef cattle results in increased intramuscular fat in the longissimus thoracis muscle in one genotype when fed low dietary VA. Although quality grade is important for increased profitability and improving taste characteristics of beef products, limiting VA too drastically can impair animal welfare. The objectives of this study were to determine if this marker-assisted management strategy would be effective, and whether any impairment in immune function would occur in a feedlot setting. Mixed breed beef steers (n=2000) were sorted into 40 feedlot pens so that all combinations of ADH1C genotype (TT or CT), VA level (50% or 100% of recommended) and hormonal implant status (implanted (IMP) or non-implanted (NI)) were equally represented within the population. The VA×ADH1C interaction was not observed. An implant status × ADH1C interaction was observed with average daily gain (ADG; P=0.03). Steers that were IMP and CT had higher ADG than IMP TT (CT=1.69 and TT=1.62 kg/day), whereas both genotypes in the NI steers were lower (CT=1.29 and TT=1.32 kg/day). Implant status was shown to affect dry matter intake (DMI; IMP=8.55 and NI=7.87 kg; P<0.01), total days-on-feed (IMP=164.4 and NI 210.5 days; P<0.01), USDA yield grade (YIELD; IMP=2.40 and NI=2.77; P<0.01), marbling score (MARB; IMP=392 and NI=455; P<0.01), longissimus thoracis area (LTA; IMP=85.0 and NI=80.7 cm2; P=0.01) and backfat thickness (FAT; IMP=8.0 and NI 10.0 mm; P<0.01). Overall, IMP animals finished on fewer total days-on-feed with higher ADG, DMI, larger LTA, and lower YIELD, MARB and FAT. To investigate immune function parameters, crossbred steers (n=18) were selected from a prior feeding trial so that all combinations of ADH1C (TT, CT and CC) and VA (25% or 75%) were equally represented. Blood cell count analysis and peripheral blood mononuclear cell proliferation and stimulation assays were conducted. None of these immune parameters were affected by VA level. Treatment and mortality records were examined in the 2000 steer population, where no correlations with ADH1C, implant status or VA level were observed. Due to no VA × ADH1C interaction, this nutrigenetic marker-assisted management strategy is not effective at this time in commercial beef cattle feedlots, however, supplementing VA at a level as low as 25% of recommended in finishing rations would likely not result in signs of immune dysfunction.

Correction to: Process-induced degradation of bioresorbable PDLGA in bone tissue scaffold production.

The original version of this article unfortunately contained a mistake. E. Themistou was missing from the author group and so is now included with this erratum.

Process-induced degradation of bioresorbable PDLGA in bone tissue scaffold production.

Process-induced degradation of clinically relevant resorbable polymers was investigated for two thermal techniques, filament extrusion followed by fused deposition modelling (FDM). The aim was to develop a clear understanding of the relationship between temperature, processing time and resultant process-induced degradation. This acts to address the current knowledge gap in studies involving thermal processing of resorbable polymers. Poly(DL-lactide-co-glycolide) (PDLGA) was chosen for its clinically relevant resorption properties. Furthermore, a comparative study of controlled thermal exposure was conducted through compression moulding PDLGA at a selected range of temperatures (150-225 °C) and times (0.5-20 min). Differential scanning calorimetry (DSC) and gel permeation chromatography (GPC) were used to characterise thermally induced degradation behaviour. DSC proved insensitive to degradation effects, whereas GPC demonstrated distinct reductions in molecular weight allowing for the quantification of degradation. A near-exponential pattern of degradation was identified. Through the application of statistical chain scission equations, a predictive plot of theoretical degradation was created. Thermal degradation was found to have a significant effect on the molecular weight with a reduction of up to 96% experienced in the controlled processing study. The proposed empirical model may assist prediction of changes in molecular weight, however, accuracy limitations are highlighted for twin-screw extrusion, accredited to high-shear mixing. The results from this study highlight the process sensitivity of PDLGA and proposes a methodology for quantification and prediction, which contributes to efforts in understanding the influence of manufacture on performance of degradable medical implants.

SAR and characterization of non-substrate isoindoline urea inhibitors of nicotinamide phosphoribosyltransferase (NAMPT).

Herein we disclose SAR studies that led to a series of isoindoline ureas which we recently reported were first-in-class, non-substrate nicotinamide phosphoribosyltransferase (NAMPT) inhibitors. Modification of the isoindoline and/or the terminal functionality of screening hit 5 provided inhibitors such as 52 and 58 with nanomolar antiproliferative activity and preclinical pharmacokinetics properties which enabled potent antitumor activity when dosed orally in mouse xenograft models. X-ray crystal structures of two inhibitors bound in the NAMPT active-site are discussed.

Discovery and Characterization of Novel Nonsubstrate and Substrate NAMPT Inhibitors.

Cancer cells are highly reliant on NAD+-dependent processes, including glucose metabolism, calcium signaling, DNA repair, and regulation of gene expression. Nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme for NAD+ salvage from nicotinamide, has been investigated as a target for anticancer therapy. Known NAMPT inhibitors with potent cell activity are composed of a nitrogen-containing aromatic group, which is phosphoribosylated by the enzyme. Here, we identified two novel types of NAM-competitive NAMPT inhibitors, only one of which contains a modifiable, aromatic nitrogen that could be a phosphoribosyl acceptor. Both types of compound effectively deplete cellular NAD+, and subsequently ATP, and produce cell death when NAMPT is inhibited in cultured cells for more than 48 hours. Careful characterization of the kinetics of NAMPT inhibition in vivo allowed us to optimize dosing to produce sufficient NAD+ depletion over time that resulted in efficacy in an HCT116 xenograft model. Our data demonstrate that direct phosphoribosylation of competitive inhibitors by the NAMPT enzyme is not required for potent in vitro cellular activity or in vivo antitumor efficacy. Mol Cancer Ther; 16(7); 1236-45. ©2017 AACR.

Methylpyrrole inhibitors of BET bromodomains.

An NMR fragment screen for binders to the bromodomains of BRD4 identified 2-methyl-3-ketopyrroles 1 and 2. Elaboration of these fragments guided by structure-based design provided lead molecules with significant activity in a mouse tumor model. Further modifications to the methylpyrrole core provided compounds with improved properties and enhanced activity in a mouse model of multiple myeloma.

Interaction of vitamin A supplementation level with ADH1C genotype on intramuscular fat in beef steers.

Previously, the single nucleotide polymorphism in alcohol dehydrogenase (ADH1C c.-64T>C) was shown to have an association with intramuscular fat (IMF) in the longissimus thoracis (LT) muscle when vitamin A was limited in finishing rations of beef steers. The purpose of this study was to determine the optimum vitamin A supplementation level, in combination with ADH1C genotype, to increase IMF of the LT muscle. In total, 45 TT genotype, 45 CT and 27 CC Black Angus crossbred steers were backgrounded on a commercial ration containing 3360 IU vitamin A/kg dry matter (DM). During finishing, the steers were randomly assigned to one of three vitamin A treatments at 25%, 50% and 75% of the National Research Council recommendation of 2200 IU/kg DM. Treatments were administered via an oral bolus. Carcass quality was evaluated and a sample from the LT muscle was collected for analysis of IMF. A treatment×genotype interaction (P=0.04) was observed for IMF; TT steers on the 75% treatment had higher IMF relative to CT and CC steers on the same treatment. Western blot analysis showed that TT steers had higher (P=0.02) ADH1C protein expression in hepatic tissue. Previously, TT steers exhibited increased IMF when fed limited vitamin A. In the current study, the lack of variation in IMF between treatments and genotypes at the lower vitamin A treatment levels was likely due to the majority of the steers grading Canada AAA (USDA Choice). However, the western blot data supports that TT steers are expected to have higher IMF deposition, due to an increased production of ADH1C. The interaction between ADH1C genotype and vitamin A supplementation level has the potential for use in marker-assisted management programs to target niche markets based on increased marbling.

Osteogenic cell response to 3-D hydroxyapatite scaffolds developed via replication of natural marine sponges.

Bone tissue engineering may provide an alternative to autograft, however scaffold optimisation is required to maximize bone ingrowth. In designing scaffolds, pore architecture is important and there is evidence that cells prefer a degree of non-uniformity. The aim of this study was to compare scaffolds derived from a natural porous marine sponge (Spongia agaricina) with unique architecture to those derived from a synthetic polyurethane foam. Hydroxyapatite scaffolds of 1 cm(3) were prepared via ceramic infiltration of a marine sponge and a polyurethane (PU) foam. Human foetal osteoblasts (hFOB) were seeded at 1 × 10(5) cells/scaffold for up to 14 days. Cytotoxicity, cell number, morphology and differentiation were investigated. PU-derived scaffolds had 84-91% porosity and 99.99% pore interconnectivity. In comparison marine sponge-derived scaffolds had 56-61% porosity and 99.9% pore interconnectivity. hFOB studies showed that a greater number of cells were found on marine sponge-derived scaffolds at than on the PU scaffold but there was no significant difference in cell differentiation. X-ray diffraction and inductively coupled plasma mass spectrometry showed that Si ions were released from the marine-derived scaffold. In summary, three dimensional porous constructs have been manufactured that support cell attachment, proliferation and differentiation but significantly more cells were seen on marine-derived scaffolds. This could be due both to the chemistry and pore architecture of the scaffolds with an additional biological stimulus from presence of Si ions. Further in vivo tests in orthotopic models are required but this marine-derived scaffold shows promise for applications in bone tissue engineering.

The effect of hair colour on anaesthetic requirements and recovery time after surgery.

Patients with red hair are much more likely to have a variant of the melanocortin-1 receptor gene and this may affect sensitivity to general anaesthetics and pain response. We did a prospective, matched cohort study of 468 healthy adult patients undergoing general anaesthesia for elective surgery. All patients received an inhalational general anaesthetic. Anaesthetic drugs and doses used, hypnotic depth, recovery times, pain scores and quality of recovery scores were recorded. More men than women had red hair, so we did subgroup and multivariable analyses to account for this imbalance. There was no significant difference in recovery times, pain scores or quality of recovery scores in those with red hair. After adjusting for age, sex, American Society of Anesthesiologists physical status and duration of surgery, the recovery ratio for time to eye-opening in redheads was comparable to those with black or brown hair, 0.82 (0.57-1.19), P=0.30. We found no evidence that patient hair colour affects anaesthetic requirements or recovery characteristics in a broad range of surgical procedures.

Genetic diversity and admixture among Canadian, Mountain and Moorland and Nordic pony populations.

As part of the requirements of the Convention on Biological Diversity, Canada has been investigating the genetic diversity of its native equine and pony populations. Along with examining four indigenous Canadian equine populations (Canadian horse, Lac La Croix pony, Newfoundland pony and Sable Island population), another 10 Mountain and Moorland, three Nordic, four horse and two feral equine populations (thought to have influenced some pony breeds) were also investigated. In total, 821 individuals were genotyped at 38 microsatellite loci. Results of the analysis of molecular variance indicated that 13.3% of genetic diversity was explained by breed differences, whereas 84.6% and 2.1% of diversity came from within and among individuals, respectively. The average effective number of alleles and allelic richness was the lowest in the Eriskay (2.51 and 3.98) and Lac La Croix (2.83 and 4.01) populations, whereas it was highest in the New Forest (4.31 and 6.01) and Welsh (4.33 and 5.87) breeds, followed closely by the Newfoundland-CDN (4.23 and 5.86) population. Expected heterozygosities varied from 0.61 in the Lac La Croix to 0.74 in the Welsh and in Newfoundland. Observed heterozygosities ranged from 0.57 in the Exmoor and 0.58 in the Sable Island herd to 0.77 in the Kerry Bog and 0.76 in the New Forest breeds. Structure and admixture analyses revealed that the most likely number of clusters was 21, although some substructure was also observed when K = 16, compared with the 24 predefined populations. Information gathered from this study should be combined with other available phenotypic and pedigree data to develop, or amend, a suitable conservation strategy for all populations examined.

The impact of vitamin A restriction and ADH1C genotype on marbling in feedlot steers.

A novel SNP was discovered within the promoter region of alcohol dehydrogenase 1C (ADH1C c.-64T>C), the C allele eliminating a potential binding site for the transcription factor C/EPBα. The purpose of this study was to examine if an interaction between this SNP and vitamin A restriction had an effect on carcass characteristics in beef cattle. Following backgrounding on a ß-carotene-deficient diet, 130 steers (50 TT, 50 CT, and 30 CC) were finished for 5 mo and received either no supplemental vitamin A (unsupplemented) or 750,000 IU/mo (supplemented). A subgroup of 5 steers • genotype(-1) • treatment(-1) was randomly selected for pre- and postfinishing liver biopsies to assess vitamin A status and measure gene expression. Unsupplemented steers (Bos taurus) had significantly greater (P < 0.05) marbling scores than supplemented steers. There was a significant interaction between genotype and vitamin A supplementation on ether-extractable intramuscular fat (IMF). Within the unsupplemented treatment, TT steers had nearly 23% greater IMF than CC steers. Additionally, unsupplemented TT steers had over 24% greater IMF than supplemented TT steers. Expression of ADH1C in the liver was additive with each additional T allele, potentially due to the elimination of a possible binding site for C/EBPα. It is plausible that CC cattle have reduced ability to metabolize retinol to retinaldehyde (and subsequently retinoic acid) and that a phenotypic effect is only observed when vitamin A is limiting. Therefore, ADH1C c.-64T>C genotype, in combination with reduced vitamin A supplementation, could potentially be implemented in marker-assisted management to maximize marbling in finishing cattle.

A study of the relationship between process conditions and mechanical strength of mineralized red algae in the preparation of a marine-derived bone void filler.

Bone void fillers that can enhance biological function to augment skeletal repair have significant therapeutic potential in bone replacement surgery. This work focuses on the development of a unique microporous (0.5-10 microm) marine-derived calcium phosphate bioceramic granule. It was prepared from Corallina officinalis, a mineralized red alga, using a novel manufacturing process. This involved thermal processing, followed by a low pressure-temperature chemical synthesis reaction. The study found that the ability to maintain the unique algal morphology was dependent on the thermal processing conditions. This study investigates the effect of thermal heat treatment on the physiochemical properties of the alga. Thermogravimetric analysis was used to monitor its thermal decomposition. The resultant thermograms indicated the presence of a residual organic phase at temperatures below 500 degrees C and an irreversible solid-state phase transition from mg-rich-calcite to calcium oxide at temperatures over 850 degrees C. Algae and synthetic calcite were evaluated following heat treatment in an air-circulating furnace at temperatures ranging from 400 to 800 degrees C. The highest levels of mass loss occurred between 400-500 degrees C and 700-800 degrees C, which were attributed to the organic and carbonate decomposition respectively. The changes in mechanical strength were quantified using a simple mechanical test, which measured the bulk compressive strength of the algae. The mechanical test used may provide a useful evaluation of the compressive properties of similar bone void fillers that are in granular form. The study concluded that soak temperatures in the range of 600 to 700 degrees C provided the optimum physiochemical properties as a precursor to conversion to hydroxyapatite (HA). At these temperatures, a partial phase transition to calcium oxide occurred and the original skeletal morphology of the alga remained intact.

Effect of patient sex on general anaesthesia and recovery.

BACKGROUND: Numerous studies have shown that women emerge faster from general anaesthesia than men, and differ in their postoperative recovery profile. The extent and underlying mechanisms for these sex-related differences in general anaesthesia are unclear. METHODS: In a multicentre, prospective, matched cohort study, 500 ASA physical status I or II patients of either sex undergoing general anaesthesia for elective surgery were recruited. All subjects received a general anaesthetic through inhalation. Anaesthetic drugs and doses used, bispectral index (BIS) scores, recovery times, pain scores, and 40-item quality of recovery (QoR-40) scores for 3 days after general anaesthesia were recorded. RESULTS: Women had higher BIS scores at similar concentrations of anaesthesia (P<0.05). Time to eye-opening (P<0.01) and time to obeying commands (P<0.01) were shorter in women. Duration of recovery room stay was longer in women, who also had higher pain scores and need for treatment of nausea and vomiting (all P<0.001). QoR-40 scores for the first 3 days after general anaesthesia were lower in women (P<0.001). Plasma progesterone concentrations in women negatively correlated with the time to eye-opening (ρ=-0.53, P=0.01). CONCLUSIONS: Patient sex is an independent factor influencing the response to anaesthesia and recovery after surgery. Women emerged faster from general anaesthesia but their overall quality of recovery was poorer. Female sex hormones, particularly progesterone, might be involved, with premenopausal women having faster recovery time but poor overall recovery.

Designs from the deep: marine organisms for bone tissue engineering.

Current strategies for bone repair have accepted limitations and the search for synthetic graft materials or for scaffolds that will support ex vivo bone tissue engineering continues. Biomimetic strategies have led to the investigation of naturally occurring porous structures as templates for bone growth. The marine environment is rich in mineralizing organisms with porous structures, some of which are currently being used as bone graft materials and others that are in early stages of development. This review describes the current evidence available for these organisms, considers the relative promise of each and suggests potential future directions.

Sintering of biphasic calcium phosphates.

Biphasic calcium phosphate (BCP) discs were fabricated and then sintered using two different sintering programs to establish whether the phases present could be controlled at low and high sintering temperatures. X-ray diffraction (XRD) was used to establish the phases present after sintering and scanning electron microscopy (SEM) determined the microstructure. Sintering program 1 involved a simple heating and cooling schedule and temperatures of 1100, 1250, 1275 and 1300 degrees C. It produced samples containing an additional alpha-tricalcium phosphate (alpha-TCP) phase at temperatures above 1100 degrees C. The original ratio of hydroxyapatite/beta-tricalcium phosphate (HA/beta-TCP) could not be maintained above this temperature. Sintering program 2 combined the heating and cooling schedules of the first program with a 900 degrees C hold stage to allow alpha-TCP to beta-TCP conversion to take place. At temperatures of 1250 and 1275 degrees C, this program was successful in completely removing the alpha-TCP phase and preserving the HA:beta-TCP ratio. The SEM results show that the surface morphology of the discs was not greatly affected by choice of sintering program.

Thermal preparation of highly porous calcium phosphate bone filler derived from marine algae.

A sustainable marine-derived bioceramic with a unique porous structure has been developed for hard tissue repair. The conversion of alga was achieved through a novel technique, involving well controlled thermal processing followed by low pressure-temperature hydrothermal synthesis. In its preparation, a heat treatment step was required to remove the organic compounds from the algae, which reinforces the mineralised matrices. Its removal is necessary to prevent issue such as immune biocompatibility and ensure phase purity of the resultant biomaterial. This paper investigates the hydrothermal technique used for the transformation of mineralised red algae to hydroxyapatite that preserves the algae's unique structure. It specifically focuses on the effects of heat treatment on the morphology of the algae, TGA, SEM and hot stage XRD to quantity the changes.