Quick Assessment of the Lower Limit of Cerebral Autoregulation Using Transcranial Doppler during Cardiopulmonary Bypass in Cardiac Surgery: A Feasibility Study.

Background: During cardiac surgery, maintaining a mean arterial pressure (MAP) within the range of cerebral autoregulation (CA) may prevent postoperative morbidity. The lower limit of cerebral autoregulation (LLA) can be determined using the mean velocity index (Mx). The standard Mx is averaged over a ten second period ( Mx 10s ) while using a two second averaging period ( Mx 2s ) is faster and may record more rapid variations in LLA. The objective of this study is to compare a quick determination of LLA (qLLA) using Mx 2s with the reference LLA (rLLA) using Mx 10s . Methods: Single center, retrospective, observational study. Patients undergoing cardiac surgery with cardiopulmonary bypass. From January 2020 to April 2021, perioperative transcranial doppler measuring cerebral artery velocity was placed on cardiac surgery patients in order to correlate with continuous MAP values. Calculation of each patient's Mx was manually determined after the surgery and qLLA and rLLA were then calculated using a threshold value of Mx > 0.4. Results: 55 patients were included. qLLA was found in 78% of the cases versus 47% for rLLA. Despite a -3 mmHg mean bias, limits of agreement were large [-19 mmHg (95% CI: -13; -25), and +13 mmHg (95% CI: +6; +19)]. There was an important interobserver variability (kappa rLLA = 0.46; 95% CI: 0.24-0.66; and Kappa qLLA = 0.36; 95% CI: 0.20-0.52). Conclusions: Calculation of qLLA is feasible. However, the large limits of agreement and significant interobserver variability prevent any clinical utility or interchangeability with rLLA.

In vitro assessment of osteoblast and macrophage mobility in presence of ß-TCP particles by videomicroscopy.

ß-TCP is widely used to repair bone defects due to its good biocompatibility, macroporosity (favoring bone ingrowth) and bioresorbability. However, cell interactions with the biomaterial at the first times of implantation remain largely unknown. We have observed cell behaviors in direct contact with ß-TCP particles using long-term culture under videomicroscopy. Osteoblastlike cells (SaOs-2) and macrophages (J774.2 and mouse peritoneal macrophages) were cultured in the presence of ß-TCP particles. For each experiment, images from 20 independent fields were acquired and stored every 15 min during 8 days. At the end of the culture, they were combined to generate time lapse videos; coverslips were fixed and observed by scanning electron microscopy (SEM). SaOs-2 proliferation was determined by counting cells on six different and independent fields at days 1, 3, and 6. Videos showed the capacity of cells to displace the particles. Dynamic follow-up showed active proliferation of SaOs-2 occurring in the direction of the particles. J774.2 and peritoneal macrophages did not proliferate but came in direct contact with the particles and actively eroded them. SEM showed that cells were stretched and fixed onto the surface and seemed to climb from the coverslip to the particles. The long-term culture under videomicroscopy allowed a better understanding of the colonization process of ß-TCP particles by osteoblastlike cells and macrophages. Data obtained from long-term videomicroscopy are in agreement with in vivo observations confirming the interest of ß-TCP to promote osteogenesis.

In vitro calcification of chemically functionalized carbon nanotubes.

Bone is composed of two phases. The organic phase is made of collagen fibrils assembled in broad fibers acting as a template for mineralization. The mineral phase comprises hydroxyapatite (HAP) crystals grown between and inside the collagen fibers. We have developed a biomimetic material using functionalized carbon nanotubes as scaffold to initiate in vitro mineralization. Biomimetic formation of HAP was performed on single-walled carbon nanotubes (SWCNTs) which have been grafted with carboxylic groups. Two types of nanotubes, HiPco(R) and Carbon Solutions(R), were oxidized via various acidic processes, leading to five different groups of carboxylated nanotubes, fully characterized by physical methods (thermogravimetric analysis, attenuated total reflectance infrared spectroscopy and X-ray photoelectron spectroscopy). All samples were dispersed in ultra-pure water and incubated for 2weeks in a synthetic body fluid, in order to induce the calcification of the SWCNTs. Scanning electron microscopy (SEM) and energy-dispersive X-ray analysis studies showed that Ca(2+) and PO(4)(3-) ions were deposited as round-shaped nodules (calcospherites) on the carboxylated SWCNTs. Fourier transform infrared and Raman spectroscopic studies confirmed the HAP formation, and image analysis made on SEM pictures showed that calcospherites and carboxylated SWCNTs were packed together. The size of calcospherites thus obtained in vitro from the HiPco(R) series was close to that issued from calcospherites observed in vivo. Functionalization of SWCNTs with carboxylic groups confers the capacity to induce calcification similar to woven bone.

In vitro kinetic study of growth and mineralization of osteoblast-like cells (Saos-2) on titanium surface coated with a RGD functionalized bisphosphonate.

Osteoconduction and osseointegration are the critical stages for implantation success. Peptides containing RGD (Arg-Gly-Asp) adhesive sequence are known to promote cell adhesion and consequently to favor osseointegration of medical devices. In this study, RGD peptides were coupled to a bisphosphonate used as an anchor system and chemically adsorbed on polished titanium discs. Two different concentrations, 10(-10) mol/L (RGD 10(-10)) and 10(-4) mol/L (RGD 10(-4)) were compared to non coated discs (RGD 0). Adhesion, spreading, and mineralization of osteoblast-like cells (Saos-2) were assessed. Mineralization kinetic was done at 3, 6, 10, 14, and 18 days of culture; the extent of mineral deposits was quantified by image analysis. Histogram repartitions of nuclear area, characterizing cell spreading, showed a shift to higher values in cells cultured on RGD coated titanium disks. Mineralization started at day 3 in the three groups, but had a faster development in the RGD 10(-10) group from day 6 to day 18 compared to RGD 0 and RGD 10(-4). At day 18, the percentage of mineralized area was significantly higher for RGD 10(-10) compared to RGD 0 (p < 0.05). In the present study, this new method was found suitable to anchor RGD containing species on titanium: this favored adhesion and spreading of osteoblast-like cells and mineralization compared to noncoated titanium.